Mycoplasma penetrans. Sequence analysis of this N-terminal region suggests the appended domain is an aminotransferase, which we demonstrate here. The aminotransferase domain of MpMetRS is capable of generating methionine from its α-keto acid analog, 2-keto-4-methylthiobutyrate (KMTB). The methionine thus produced can be subsequently attached to cognate tRNAMet in the MpMetRS aminoacylation domain. Genomic erosion in the Mycoplasma species has impaired many canonical biosynthetic pathways, causing them to rely on their host for numerous metabolites. It is still unclear if this bifunctional MetRS is a key part of pathogen life cycle or is a neutral consequence of the reductive evolution experienced by Mycoplasma species.The structural organization and functionality of aminoacyl-tRNA synthetases have been expanded through polypeptide additions to their core aminoacylation domain. We have identified a novel domain appended to the methionyl-tRNA synthetase (MetRS) of the intracellular pathogen  Aminoacyl-tRNA synthetases (AARSs) are multi-domain proteins that catalyze attachment of a specific amino acid to its cognate tRNA, preparing the aminoacyl-tRNAs for template-directed protein synthesis at the ribosome. AARSs have been partitioned into two classes based on evolutionarily conserved structure and sequence similarities in their catalytic domains ,2,3. OveMycoplasma penetrans, an obligate intracellular human pathogen first identified in HIV-infected patients . In. InMycopccupancy . The lowE. coli, we wanted to identify whether MpMetRS exists in vivo as the proposed aminotransferase-synthetase fusion protein. We therefore performed immunoblots on both M. penetrans cell lysate and recombinant proteins. The band present in each sample just below the 130 kDa marker is consistent with the 126.4 kDa size of full-length MpMetRS  was performed to assess the molecular weight and polydispersity of the enzyme [As the e enzyme ,25. ThisMet synthesis [Met; the role of the MpMetRS appended domain was unclear. Consistent with our earlier findings, aminoacylation conducted in this work, using full-length MpMetRS (M568A), remained robust for its tRNA substrate  in the aminotransferase domain; this conserved lysine aligns with the Lys-209 residue of transfer . Two indnticodon ,27,28. Te enzyme .32P-labeled tRNAMet. When MpMetRS is supplied with methionine directly, Met-tRNAMet is formed, as previously shown. When the aminotransferase is supplied with phenylalanine and KMTB in the absence of methionine, Met-tRNAMet is synthesized, albeit at reduced efficiency . Aminoacylation from KMTB alone is negligible (0.04 pmol/min). The reaction kinetics of each domain were determined through either LC–MS or aminoacylation assays and are summarized in The ability of MpMetRS to use KMTB as a transaminase substrate and the presence of fused aminotransferase and tRNA synthetase active sites led us to investigate whether the product of one domain could provide the substrate for the other. To test this hypothesis, we performed an aminoacylation assay in the presence of KMTB and phenylalanine with ficiency . FormatiMycoplasma spp. and is to our knowledge unique to M. penetrans MetRS. While many AARSs contain appended domains that expand their functionality beyond tRNA aminoacylation , MpMetRSM. penetrans cell lysates confirmed that the full-length protein does exist in vivo. The presence of both full-length and core MpMetRS suggests that both forms are biologically relevant, although the larger polypeptide is more abundant in the cell lysates analyzed here. It remains to be seen whether the ratio of full-length to core proteins varies with cellular conditions. The likelihood that an internal ribosomal start site is responsible for the two alternate forms upon overexpression in E. coli is strengthened by observation of a single polypeptide product in the M568A variant corresponding to full-length protein  were used in an aminoacylation reaction, MpMetRS and M568A aminoacylate Mp-tRNA enzymes .M values for the Mp tRNAMet substrate, there is a notable difference in the chemical step of aminoacylation, represented by kcat  corresponds to a structural switch from dimer to monomer. The SEC–MALS analysis for MpMetRS∆ suggests a molecular weight of 77 kDa, somewhat higher than the calculated 61.4 kDa. The MpMetRS∆ sample did display greater scatter in generated molecular weights , suggest by kcat . While e by kcat , structuThe aminotransferase domain of MpMetRS was shown to utilize more than one set of amine group donor and acceptor. While some aminotransferase enzymes are specific for a given amino acid-keto acid pair, others are more promiscuous in their substrate use ,30,31,32Met. Over the course of two minutes, the initial rate for Met-tRNAMet synthesis beginning with KMTB and Phe was half that observed with methionine added directly to the reaction (We have shown here that the fused aminotransferase domain is capable of producing methionine that can be used by the core synthetase domain to synthesize Met-tRNAreaction . For theE. coli MetRS (PDB 1QQT) and Anabaena sp. AGAT (PDB 1VJO) [3 group from aqueous environment through a hydrophobic tunnel to the PRPP active site 16 Å away [Met, while preventing enzymes like S-adenosylmethionine synthase from competing for its common substrate. The MpMetRS fusion protein could allow direct transfer of methionine from the aminotransferase domain to the aminoacylation domain, either within one monomer (intramolecular channeling) or across the dimer interface (intermolecular channeling). Alternatively, methionine may simply be released from the aminotransferase active site for use by a neighboring enzyme. While the existence of a bifunctional enzyme does not necessitate substrate channeling, there is precedent for monofunctional enzymes that exist as a fusion protein in another organism. One clear example is the thymidylate synthase (TS) and dihydrofolate reductase (DHFR) enzyme pair; in humans, they are isolated monofunctional enzymes, while in some protozoan species, they are bifunctional fusion proteins [kcat ~6 s−1). Additionally, there was little benefit to providing the enzyme time to produce methionine prior to tRNA addition, suggesting that simple diffusion may not explain how the MetRS sequesters its methionine substrate. Future work includes structural investigations of the individual catalytic domains and the full protein fusion to understand whether substrate channeling is possible.Given the two distinct active sites on this novel enzyme, and the capacity for the synthetase domain to use methionine produced by the aminotransferase domain, the question of methionine transport arises. Crystal structures exist for isolated proteins homologous to these domains: DB 1VJO) ,34. Howe6 Å away . A ratio6 Å away . One evoproteins . Substraproteins . Our kinM. penetrans would retain such a gene fusion in the context of a condensed genome lacking many key metabolic enzymes [M. penetrans lacks methionine biosynthesis enzymes. Use of an aminotransferase would enable M. penetrans to increase the local concentration of methionine, using KMTB and an amine-donating amino acid. Synthesis of methionine from KMTB is the last step in the methionine salvage pathway; our data suggest that the N-terminal domain of MpMetRS is a transaminase for methionine production.The question remains as to why  enzymes ,40. As tTreponema denticola [Anabaena spp. AGAT. It remains to be determined if the MpMetRS aminotransferase has an affinity for 2-aminoethylphosphonates or whether it possesses nucleotidyltransferase capabilities suggested by this new alignment. Future work will focus on structure determination of this novel fusion protein and its possible role in the metabolic life cycle of the human pathogen M. penetrans.Recombination events during genomic erosion of mycoplasma species may have led to unique protein fusions like the one seen with MpMetRS. The essentiality of the MpMetRS appended domain remains unknown, despite the demonstration here that it possesses aminotransferase activity in vitro. Until recently, the extreme N-terminal 166 amino acids of MpMetRS did not align with any known proteins; however, a PHYRE2 structure-based search of the MpMetRS appended domain suggested homology to another fusion protein, phosphonate-specific cytidylyltransferase, and 2-aminoethylphosphonate transaminase (PDB 6PD1), from the bacterium enticola . The 2-aM. penetrans life cycle and whether the acquisition of the aminotransferase domain is unique among AARS enzymes or may yet be identified in additional species.In conclusion, the results presented here demonstrate the first example of an AARS for which the canonical aminoacylation activity is augmented by aminotransferase function. The capacity for MpMetRS to generate its own methionine substrate from a metabolic precursor suggests an intriguing adaptation to environmental conditions. It remains to be determined whether this catalytic duality contributes to the


The adverse effects of fat mass on functional dependence might be attenuated or worsened, depending on the level of muscular strength. The aim of this study was to determine (i) the detrimental effect of excess adiposity on dependence in activities of daily living (ADL), and (ii) whether relative handgrip strength (HGS) moderates the adverse effect of excess adiposity on dependence, and to provide the threshold of relative HGS from which the adverse effect could be improved or worsened. A total of 4169 participants (69.3 ± 7.0 years old) from 244 municipalities were selected following a multistage area probability sampling design. Measurements included anthropometric/adiposity markers ), HGS, sarcopenia “proxy” , and ADL . Moderation analyses were performed to identify associations between the independent variable (WHtR) and outcomes (dependence), as well as to determine whether relative HGS moderates the relationship between excess adiposity and dependence. The present study demonstrated that (i) the adverse effect of having a higher WHtR level on dependence in ADL was moderated by relative HGS, and (ii) two moderation thresholds of relative HGS were estimated: 0.35, below which the adverse effect of WHtR levels on dependency is aggravated, and 0.62, above which the adverse effect of fat on dependency could be improved. Because muscular strength represents a critically important and modifiable predictor of ADL, and the increase in adiposity is inherent in aging, our results underscore the importance of an optimal level of relative HGS in the older adult population. Muscle strength and mass decline with aging . The impSeveral studies have indicated that the aging process produces a series of changes in body composition, usually without affecting the body mass index or weight, but producing an accumulation of fat as individuals get older ,7. BeyonHandgrip strength (HGS) is the most common index of muscle strength, owing to its ease of assessment, low cost, and simplicity, and it is considered a valid “proxy” of overall muscle strength for clinical and epidemiological studies . Lower HThis worsening of muscle strength can be explained several biological factors. First, fat infiltration induces changes in contractile function  in the dThe adverse effects of abdominal obesity on functional dependence, might be attenuated or worsened depending on the level of muscular strength. Additionally, relative handgrip strength is associated with functional dependence. Thus, central adiposity may have an effect between dependence status and relative handgrip strength after potential confounding variables such as age, gender, and/or lifestyle. Accordingly, describing the magnitude of these risk factors in older adults could be important for prioritizing prevention and public health efforts. Nevertheless, to our knowledge, no studies have examined the moderator role of muscle strength based on HGS between excess of central adiposity and functional dependence.The aim of the present study was two-fold: (i) to examine the detrimental effect of abdominal obesity on functional dependence in ADL, and (ii) to discern whether relative HGS moderates the adverse effect of abdominal obesity on dependence, as well as to provide the threshold of relative HGS from which the adverse effect could be improved or worsened.The data for this secondary cross-sectional study were obtained from the Health and Well-being and Aging Survey in Colombia 2015 , a multicenter project conducted from 2014 to 2015 by  . The stuA total of 23,694 surveys were conducted at the national level. A total of 6530 segments were planned to obtain the surveys , with an expected average of 4.7 adults per segment. The standardized process for each home visit involved the identification of the participants, the registration of the demographic data, the signing of the informed consent, the application of the established filters and the selection criteria, the signing of assent when necessary, and the completion of the questionnaire by the interviewer. For this subsample, the calculation of the sample size was carried out, taking into account national representation. A total of 86 municipalities were selected, including the four large cities. For this analysis, we used data from 4169 participants included as a subsample with HGS measures. The rationale and detailed methodology of the SABE Colombia has been described in another document [Institutional review boards involved in developing the SABE 2015 study  reviewed and approved the study protocol. The study protocol for the secondary analysis was approved by the Human Subjects Committee at the Pontificia Universidad Javeriana  in accordance with the Declaration of Helsinki of the World Medical Association and Resolution 8430 of 1993 of the then Ministry of Health of Colombia on technical, scientific, and administrative standards for conducting research with humans. All participants provided written informed consent.Data collection staff were trained by the research teams of the coordinating centers  for face-to-face interviews and physical measurements. Anthropometry measurements included height and body weight, which were measured with a portable stadiometer  and an electronic scale , respectively. Body mass index (BMI) was calculated in kilograms per square meter from the measured body weight and height. Waist circumference (WC) was measured over the midpoint between the lower border of the ribs and iliac crest in the midaxillary plane, at the end of normal expiration. The waist-to-height ratio (WHtR) was measured as the ratio of the waist circumference (in cm) to the height (in cm). We used WHtR as a measure for abdominal obesity because is a useful tool in clinical practice and has been shown to be a reliable parameter for predicting whole-body fat percentage and visceral adipose tissue . The calNutritional status was evaluated through Mini-Nutritional Assessment extended version . FunctioFor lifestyle characteristics, personal habits regarding alcohol intake  and cigarette smoking  were recorded. A “proxy physical activity” report was conducted by the following questions: (i) “Have you regularly exercised, such as jogging or dancing, or performed rigorous physical activity at least three times a week for the past year?”; (ii) “Walk, at least three times a week, between 9 and 20 blocks (0.6 to 1.2 km) without resting?”; (iii) “Walk, at least three times a week, eight blocks (0.5 km) without resting?”. Participants were considered physically active if they responded affirmatively to two of the three questions. Medical information including multimorbidity, as well as chronic conditions adapted from the original SABE study, were assessed by asking the participants if they had been diagnosed, by a physician, with hypertension, type 2 diabetes mellitus, chronic obstructive pulmonary disease, cardiovascular diseases , stroke, cancer, arthritis, osteoporosis, or sensory impairments (vision and hearing loss). Medication use was evaluated with the question “do you currently take or use any prescription medication?”.Race/ethnicity grouped as indigenous , black “mulato” or Afro-Colombian, white, and others  was assessed by self-reporting. Socioeconomic status (SES) was determined on the basis of the housing stratum (1 to 6), with level 1 being the highest poverty and level 6 the highest wealth. This classification is a measure developed by the National Government of Colombia that considers physical characteristics of the dwellings and their surroundings. The classification in any of the six strata approximated the hierarchical socioeconomic difference from poverty to wealth and vice versa.t-test or the chi-square (χ2) post-hoc test.Descriptive analyses using mean ± standard deviation (SD) for the continuous variables and frequency distribution for categorical variables were used to obtain the characteristics of the sample. The normality of the data was examined by the Kolmogorov–Smirnoff test. Significant differences between men and women were analyzed using Student’s p = 0.814); therefore, all analyses were performed with men and women together. Moderation analysis was conducted to examine whether WHtR levels were related to increased dependence and to determine whether this negative effect was moderated by relative HGS. This relationship used ordinary least squares regression analysis when predicting continuous variables (WHtR and relative HGS in the study). A simple slope plot was used to visualize the effect of the moderator. The Johnson–Neyman approach was used to test the point in which the relative HGS value moderated the relationship between WHtR levels and dependence. The Johnson–Neyman technique determined, along a continuum of moderator values (relative HGS), the region of significance on the relationship between the independent and dependent variables [The PROCESS macro in the SPSS statistical software package, version 24.0  for Windows, was used to conduct a moderation analysis. Preliminary analysis showed no significant interactions between gender and abdominal obesity in relation to functional dependence (ariables . All tesp < 0.05), with the exception of calf circumference. Regarding performance outcomes—computed from absolute HGS and relative HGS—men showed a significantly higher performance than women. Additionally, the ethnic distribution was dissimilar between sexes, except for the Afro-Colombian ethnic group. We observed a major proportion of white and other ethnic groups . A major proportion of participants were found to be in SES level 2 and significant differences were observed between sexes in all SES levels except level 1.Of the 4169 study participants, 56.2% were female and 43.8% were male . AnthropLikewise, there were significant differences between sexes regarding lifestyle habits including smoking, alcohol consumption, and physical activity “proxy” recommendations. According to self-report comorbidities presented by participants, there was a prevalence of visual problems (57.7%) and high blood pressure (53.7%) in both sexes. Regarding the distribution difference between males and females, we found significant differences in hearing problems, high blood pressure, type 2 diabetes mellitus, arthritis, and osteoporosis. In addition, there were significant differences in medication use and nutritional status. Finally, the prevalence of dependency was 8.1% and 4.3% for mild and moderate dependency, respectively. However, when we combined all three levels of dependency  this rose to 12.5%, with 522 older adults dependent in ADL.β = −0.11 ) was moderated by relative HGS. Therefore, the adverse effect of excess adiposity on functional dependence was moderated by relative HGS ).To elucidate a possible estimate point from which the moderator value has a moderator effect, the Johnson–Neyman statistical approach was used. The result is shown in The present study investigated the moderator role of HGS on the adverse effect of WHtR on dependency in older Colombian adults. The major finding of the study was that the adverse effects of high WHtR levels on dependency were found to be moderated by relative HGS. Two moderation thresholds of relative HGS were estimated: 0.35, below which the adverse effect of WHtR levels on dependency was aggravated, and 0.62, above which the adverse effect of fat on dependency improved. Accordingly, our results indicated that older adults with higher WHtR could experience more dependence in ADL than older adults with lower WHtR; however, this unfavorable effect was moderated by relative HGS. Consequently, older adults with high relative HGS levels could attenuate the negative effect of adiposity. Therefore, age-related declines in muscle mass and strength are often detected by reductions in HGS.The findings of the present study are supported by several previous studies. For example, de Carvalho et al.  found thNo previous studies have reported a moderator role of HGS on the relationships studied here. We found that higher relative HGS could attenuate the adverse effect of abdominal obesity on functional dependence in older adults. Our findings show that higher central obesity has an adverse effect on functional independence. It is therefore likely that functional independence in ADL will be reduced in those older adults with abdominal obesity. However, this negative effect could be moderated by relative HGS. Consequently, the adverse effect may worsen, improve, or even disappear, depending on the relative HGS of older adults. Our findings indicate that muscle strength relative to body weight can play a crucial role between WHtR levels and dependency. Specifically, if older adults have a high WHtR value and a relative HGS above 0.62, the adverse effect on dependency could be mitigated or even disappear. Conversely, if older adults have a high WHtR value and a relative HGS below 0.35, it could worsen the adverse effect on dependency.Biomechanical and neuromuscular scientific evidence could justify the moderator role of HGS between excess of central adiposity and functional dependence. For example, it is well reported that abdominal obesity is related to a greater body weight, and consequently walking more slowly might help to keep the dynamic balance between steps, as well as to maintain shorter the cadence and length of steps to optimize gait pattern . AnotherThe strengths of the study include the large population-based study in older, Latin-American adults. Additionally, we carried out complex statistical analyses to determine the role of muscle strength to circumvent the detrimental effect of excess adiposity on dependence. As well as this, through the Johnson–Neyman statistical approach, we provided two thresholds of relative HGS, which we believe will add to the knowledge base to improve clinical practice and exercise programs in this population.There are some limitations of the study design that need to be considered. First, the cross-sectional design limits drawing any causal inferences. Second, the assessment of excess adiposity can result in bias because of the proxy method , and therefore, standardized measures of body composition should be used. Third, the classification of dependency was based on a self-report questionnaire. Thus, we are unable to say whether low grip strength (with or without excess adiposity) leads to higher risk of neuromuscular/ADL abnormalities, or conversely, whether poor neuromuscular/dependency profiles lead to declines in grip strength . Future research is needed to better describe the age- and sex-specific trajectories of HGS as a predictor of comorbidities across the lifespan, and perhaps, just as importantly, to apply robust analyses that can compartmentalize risk into hierarchical categories. Finally, the thresholds for HGS are open to discussion and the values may vary depending on the comorbidities that individuals present with.In summary, older adults with excess adiposity have major dependency in ADL. However, this adverse effect can be moderated by relative HGS. Our findings bring two thresholds of relative HGS as moderators of the adverse effect: <0.35, in which the adverse effect of abdominal obesity on dependence could worsen, and >0.62, in which the detrimental effect could be improved or even disappear. Because muscle strength represents a critically important and modifiable predictor of ADL ,14,43, a


Occupational related respiratory diseases arise as a result of the deposition of dust particles in the lungs. Flour milling industries; generate organic dust during industrial processes, such as cleaning, milling, packaging, and loading which release dust into the air and later inhaled by workers. Flour mill workers are at risk of developing respiratory health problems because of exposure in their working environment, but existing data were few. The aim of this study was to assess the prevalence of chronic respiratory symptoms, lung function and associated factors among flour mill factory workers.A comparative cross-sectional study was conducted among 196 flour mill factory workers and 210 soft drinks factory workers. We selected study participants using a systematic sampling technique. We assessed the chronic respiratory symptoms using the questionnaire adopted from the British Medical Research Council. Binary logistic regression analysis with 95% CI and p < 0.05 was used to identify the factors. Lung function parameters; Forced Vital Capacity (FVC), Forced Expiratory Volume in one second (FEV1) and ratio FEV1/FVC was measured by using spirometer and analyzed by using an independent t-test.We included 406 (96.7%) workers in this study. The prevalence of chronic respiratory symptoms was higher among flour mill workers as compared to soft-drinks factory workers (56.6% vs.12.9%). Primary education , mixing department , work experience 6–9 years , work experience ≥10 years  and working over eight hours  were factors that significantly associated with chronic respiratory symptoms among flour mill workers. FVC (p < 0.002), FEV1 (p < 0.001) and FEV1/FVC (p < 0.012) were significantly reduced among flour mill workers.We found chronic respiratory symptoms to be high among flour mill workers. Lower education level, mixing department, increased work experience, and longer working hours were identified factors. The flour mill dust exposed worker’s lung function parameters were highly reduced. This study suggested that workers’ dust exposure reduction and control methods in flour mill factories need to be implemented. Occupational related respiratory diseases arise because of the deposition of dust in the lungs. The deposited dust disturbs the process of inhalation and exhalation of air whose intensity is influenced by the dust, duration of exposure, concentration, and size of dust in the breathing zone and genetic factors , 2. ChroFlour is a complex organic dust containing powder finished production of milling cereals like wheat, barley, corn (maize), or a combination of these, got after several reduction processes . Flour mFlour mill workers are at risk of developing respiratory health effects due to exposure in their working environment . Lung fu1), Forced Vital Capacity (FVC) and the ratio of Forced Expiratory Volume in 1  second and Forced Vital Capacity (FEV1/FVC) among flour factory workers [Flour dust exposure causes respiratory symptoms such as chest pain, phlegm, wheezing, cough, and shortness of breathing in flour mill workers , 12–14.  workers –18.Study conducted in 1998 in Ethiopia reported that a higher prevalence of respiratory symptoms like a cough (42.1%), wheezing (35.7%) and chest tightness (35.1%) in flour mill workers but did not investigate their associated factors and lung function , 19. TheAn institutional-based comparative cross-sectional study was conducted in April 2018 among flour mill and soft drink factory workers in Hawassa city of Southern Ethiopia. In Hawassa city, there are ten flour processing factories  and one soft drink factory.One hundred ninety-six flour mill workers worked for at least one year in the factory were included in this study. In addition, we included two hundred ten soft drinks factory workers as a comparison group. We performed a lung function test only for 108 randomly selected workers (54 from the flour mill factory and 54 from the soft drink factory) from 406 study participants because of limited resources.We included all workers above 18 years of age who had direct involvement in production and working in the flour mill and soft drink factory for one year and above. Workers who had heart failure, tuberculosis, drug addicts, cigarette smokers, emphysema, recent surgery of the thorax, abdomen, undergone vigorous exercise and any acute illness were excluded from the lung function test.nd interval by taking roster of the workers in each department.First, the work process was stratified by production units in both flour mill and soft drink factories. Then, systematic sampling techniques applied to draw the required sample size. The workers’ roster was used as a sampling frame. First subject selected randomly from each department, then we selected other subjects every 2Chronic respiratory symptoms: are defined as the development of one or more of the respiratory symptom(s) of a chronic cough, chronic cough with sputum, chronic breathlessness, chronic wheezing, chronic chest illness which lasts at least three months in one year , 21.1)  and FEV1/FVC; the percentage of the FVC expired in the first second of maximal forced expiration following full inspiration [Lung function reduction: Forced vital capacity (FVC); the maximum volume of air that can be breathed out forcefully and rapidly following a maximum inspiration, Forced expiratory volume in 1 second , sex , educational status  and monthly income (20 US$-70 US$ and ≥ 70 US$) were included.(carrying of flour from the packing areas to the stores or trucks).The behavioral factors of workers (like a smoking habit); current smokers (workers who were smoking at the time of the study or a person who smoke cigarettes every day or some days) and every smoker: a worker who has smoked at least one hundred cigarettes during his/her life, which includes current smokers . The worPrevious exposure history; workers experience in the dusty environment before the current working position. Previous medical history  that could be developed before and confirmed by physicians.We conducted the study after having an ethical clearance from the Institutional Review Board of the College of Health Sciences of Addis Ababa University. Before performing measurements, we obtained verbal and written consent from each study participant and participants informed that they have full rights to refuse and discontinue taking part at any point in the study. The study participants with lung function impairments were advised and linked to a health facility.Data were collected by using questionnaires adopted from British Medical Research Council (BMRC) . The comWeight of study participants were measured by using a standardized electronic weighing machine, with the subjects standing and wearing light clothes and height of the subjects measured with the stadiometer with portable field survey scales. Body mass index (BMI) calculated by using Ndd Medical technologies’ software.Ndd Easy on-PC spirometer was used for determining lung function parameters  among both flour mill and soft drink factory workers according to the American thoracic society recommendation . Before We performed all measurements in the sitting position. While doing this maneuver, flow and volume curves inspected on the screen for detecting whether subjects displayed enough effort during inspiration and expiration. Repeatability of FEV1 and FVC parameters were checked and three acceptable maneuvers with repeatable results were taken and we recorded the highest reading of these. Predicted values for lung function tests were not obtained due to lack of reference equation for the Ethiopian population. Trained and certified health professionals selected from different hospitals performed lung function tests. Standard operative procedures (SOPs) were used to improve quality of lung function test. In addition, the volume was measured by using 3 l syringe at the beginning of the test.Collected data were organized and entered Epi data version 3.1 and we did cleaning to avoid missing values, outliers, and other inconsistencies. For data cleaning, frequency, sort, and list were used. Cleaned data exported to SPSS version 24 for analysis. Descriptive statistics were used to summarize data. Prevalence odds ratio with 95% CI was used to compare the prevalence of chronic respiratory symptoms of a flour mill with soft drink factory workers. Logistic regression analysis used to identify whether exposure variables are significantly associated with outcome variables or not. Thus, variables with p < 0.2 were included in the multivariable analysis by adjusting confounding variables; age, smoking habit, income, educational status, past dust exposure, working hours per day. The P-value of < 0.05 was considered as statistically significant. An independent t-test was used for continuous data; to compare the lung function test between flour mill and soft drink factory workers.Four hundred six (96.7%) workers were included in this study , from 10 flour mill factories and one soft drink factory in Hawassa city, southern Ethiopia. From 420 flour mill workers, 4 workers were refused to take part, 4 workers were on sick leave, and six had stopped working in the factory. Most study participants were males, which accounts for 78.3% and (mean ± SD) age of flour mill workers were 32.1 ± 9.99 and for a soft drink, factory workers were 31.28 ± 7.87 and ranging from 20 to 63 years. Seventy-three (73%) of participants from flour mill factories had attained a primary level of education and about 103 (49%) of soft drink factory participants had attained more than secondary education.One hundred sixty-three (83.2%) of participants from flour mill factories had a monthly income between 20 US$ and 70 US$ and, 167 (79.5%) of soft drink factory participants ≥70 US$. There was a significant difference between the flour mill factories and soft-drinks factory participants in terms of age, educational status, income and smoking habit (ever smoker) (p < 0.05). In this study, 39 (19.9%) of flour mill factory workers and 4 (1.9%) of soft-drinks factory workers were ever smokers (Table Hundred one (51.5%) of flour mill factory workers and 24 (14.4%) of soft drink factory workers had an experience of dust exposure before joining the present job. From flour mill factory workers about 34% had work experience in the factory for six years or more and about 36% had work experience for over ten years. One hundred forty-five (69%) of the soft-drinks factory and 95 (46.5%) of flour mill factory workers were working greater than 8 h per day. Few flour mill factory workers 11 (5.6%) and soft drink factory workers 9 (4.3%) were reported the chronic respiratory disease confirmed by physicians before they have started a job in the factory. There was a significant difference between flour mill factory workers and soft-drinks factory workers in terms of duration of working hours and past dust exposure, p < 0.05 , chronic cough with sputum , chronic wheezing , chronic breathlessness  and at least one chronic respiratory symptoms (56.6% Vs. 12.86%). The odds of chronic respiratory symptoms in flour mills are 32% more than that of the soft drink factory  . Workers who were engaged in mixing department were 5.3 times more likely to develop chronic respiratory symptoms than workers engaged in cleaning, loading and packaging sections . Workers who had work experience 6–9 years  and work experience ≥10 years  had the odds of developing chronic respiratory symptoms 5.1 and 2.5 times more than workers who had work experiences between 1 and 5 years, respectively. Workers who stay greater than 8 h per day were 2.4 times more likely to develop chronic respiratory symptoms than those who had 8 h or less .Table We included one hundred eight male workers (54 from the flour mill factory and 54 from the soft drink factory). Six Spirometers results among flour mill workers and six among the soft-drinks factory workers were excluded from the analysis because of unacceptable readings. We present the mean values of the lung function parameters for the total number of flour mill workers and soft in Table . The val1 (p < 0.001) and FEV1/FVC (p < 0.012).The prevalence of almost all chronic respiratory symptoms was higher among flour mill factory workers, compared with soft-drinks factory workers. Educational status, mixing department, working hours and service years of workers were statistically significantly associated with chronic respiratory symptoms. In this study, flour mill workers showed a statistically significant reduction in FVC (p < 0.002), FEVThe development of at least one chronic respiratory symptom was higher among flour mill workers (56.6%) compared to the soft-drinks factory (12.86%). This study showed higher prevalence of chronic respiratory symptoms than in other flour mill factory studies, such as those in Ibadan, Nigeria (54% of production staff and 19% in controls), in Ilorin, North Central Nigeria (49.5% of the flour mill workers and 27.7% in controls) , 27.The most common chronic respiratory symptoms in the study were chronic cough 39.3%, chronic breathlessness 18.9%, chronic cough with sputum 17.86% and chronic wheezing 17.35%. The result was higher than the study done in Addis Ababa, Ethiopia in flour mill workers, which was 27.7% dry cough, 14.8% wheeze, and 16.6% breathlessness [On the other hand, the result of this study was lower than the study conducted in flour mill factories in Egypt that reported the prevalence of cough 87.5%, shortness of breath 60% and wheezes 72.5% [There is inadequate personal protective equipment in the work environment and the workers did not wear the personal protective equipment properly even if little equipment is available. This study finding is in line with studies conducted in India and Nigeria flour mill workers found that the most of the workers didn’t use personal protective equipment while working , 15, 28.In this study, primary school educated workers were more likely at risk of developing chronic respiratory symptoms than workers who got secondary education and above. This result was in line with a study conducted in the Ethiopian factory . Low eduIn this study, mixing department workers were more likely to develop chronic respiratory symptoms than workers in packaging, loading and cleaning departments. This finding was inconsistent with studies conducted in Iraq and Egypt, which reported that packaging department workers had a high prevalence of chronic respiratory symptoms compared to those of mixing, loading and cleaning department’s workers , 30. ThiThis study showed that workers who had work experience 6–9 years and ≥ 10 years had the odds of developing chronic respiratory symptoms were 5.1 and 2.5 times more than workers who had work experiences between 1 and 5 years, respectively. This finding was in line with the studies conducted in Iraq and Egypt flour mill workers , 30. Thi1 and FEV1/FVC compared to the soft-drinks factory workers. Meo et al., Dhillon et al., Meo and AL-drees and Melo et al. reported that flour mill workers had significantly reduced lung function parameters FVC and FEV1 [1, and FEV1/FVC were significantly reduced among flour mill workers. This might be due to the difference in working conditions such as ventilation type, machine type and use of the protective device.The present study’s results showed that among flour mill dust exposed workers a significant decline in the lung function parameters, FVC, FEVand FEV1 , 15, 31.1 and FEV1/FVC than the control group [1 and FEV1/FVC in the bakery workers were significantly lower than those of the control subjects [1 and FEV1/FVC among the flour mill workers might be due to increased dust accumulation in the respiratory system associated with prolonged exposure at workplaces.These study results were consistent with studies conducted by Mohammadien et al. and Deshpande et al., who observed that lung function parameters reduced in non-smoking flour processing workers and reported that the exposed group had significantly lower FVC, FEVol group , 32. In subjects . The redOne of the limitations of this study is healthy workers effect; workers who developed the symptoms may have quitted the job. The use of questionnaire method was also another limitation of this study because it may cause participants recall bias and interviewer bias. In addition, it would have been helpful if job designations for all workers were recorded and described in results since there may be specific types of workers at greater risk of respiratory symptoms. Furthermore, spirometry was not performed on all participants post-bronchodilator was not done in this study.Chronic respiratory symptoms were higher among flour mill factory workers than soft drink factory workers. Lower education level, increased work experience, work departments and longer working hours were determinant factors for the occurrence of chronic respiratory symptoms. The results showed that work-related exposure to a flour mill dust could reduce the mean values of lung function parameters such as FEV1, FVC, and FEV1/FVC. The study findings imply that there is a need to implement dust control methods to reduce workers’ exposure in the flour mill factory. Besides, a follow-up study with dust measurements on the effects of flour mill dust on respiratory health among flour mill production workers is recommended for the future.


Al direction, resulting in an uneven distribution of phases. Considering the flow softening degree and the excessive heterogeneous precipitation of θ′′ and θ′ phases during hot deformation, the reasonable strain rate and temperature are about 0.0003 s−1 and 150 °C, respectively. Deformation behavior and precipitation features of an Al–Cu alloy are investigated using uniaxial tensile tests at intermediate temperatures. It is found that the true stress drops with the decreased strain rate or the increased deformation temperature. The number of substructures and the degree of grain elongation decrease with the raised temperature or the decreased strain rate. At high temperatures or low strain rates, some dynamic recrystallized grains can be found. The type of precipitates is influenced by the heating process before hot tensile deformation. The content and size of precipitates increase during tensile deformation at intermediate temperatures. As the temperature increases over 200 °C, the precipitation process (Guinier Preston zone (G.P. zones)→θ′′ phase→θ′ phase) is enhanced, resulting in increased contents of θ′′ and θ′ phases. However, θ′′ and θ′ phases prefer to precipitate along the {020} In view of their excellent properties, 2xxx series Al (Al–Cu) alloys are widely used for aerospace components ,2,3,4,5.In recent years, many researches on the deformation behavior ,9,10,11 The above researches mostly concentrated on the deformation behavior and precipitation features in hot compressed Al–Cu alloys, where the deformation temperature is generally higher than 250 °C. However, few investigations focused on the tensile deformation at intermediate temperatures such as below 250 °C. Nevertheless, the researches on the deformation behavior and precipitation features in Al–Cu alloys such as 2219 aluminum alloy at intermediate temperatures are significant for the industrial production of large-scale thin-walled ellipsoidal heads. In this work, the deformation behavior of an Al–Cu alloy is investigated by uniaxial tensile tests at intermediate temperatures (100–250 °C). The changes in substructures and grains were analyzed by electron backscattered diffraction (EBSD) analysis. Furthermore, the precipitation features during tensile deformation were investigated by differential scanning calorimetry (DSC), transmission electron microscopy (TEM), and scanning electron microscopy (SEM).−1. The samples were firstly heated to experimental temperatures by 10 °C/min and then held for 5 min to acquire an even temperature field. Afterward, the samples were stretched to 15% true strain at different strain rates. Finally, the samples were cooled in air to room temperature.The initial experimental material was one typical Al–Cu alloy  rolled sheet with 12 mm thickness. Its chemical composition was 6.37 Cu–0.30 Mn–0.16 Fe–0.12 Zr–0.05 Si–0.04 Ti–Al  (wt. %). The sheet was solution-treated at 535 °C for 40 min [3 (240 mL) and CH3OH (560 mL) at −30 °C. The prepared foils were analyzed by EBSD  and TEM  detectors. Additionally, the content and type of precipitated phases were analyzed by DSC . Firstly, to acquire the baseline of DSC, an empty crucible was heated with the heating rate of 10 °C/min. Then, the samples were grinded with sandpaper and cleaned by anhydrous ethanol. The size of DSC samples was 5 mm × 5 mm × 2 mm, and the mass was between 90 and 110 mg. Next, the samples were put in a crucible, and these samples were subsequently heated with the same heating rate. To evaluate the mechanical properties of the alloy, the uniaxial tensile tests were executed on the MTS-GWT2105 test machine in the tested condition. After tensile deformation, the fracture morphologies were observed using a JEOL-7001F1 field emission (FE) SEM .To analyze the effects of deformation parameters on microstructures, foils were cut from the central part of stretched samples. The foils were polished by abrasive paper and then thinned by electro-polishing in a solution of HNO−1 and 0.0003 s−1. However, it firstly increases and then keeps stable, finally rapidly dropping with the increase in strain at 0.00003 s−1, i.e., the dynamic softening is enhanced. In −1. The true stress firstly rises and then keeps stable at 0.0003 s−1. However, obvious dynamic softening behavior is observed at 0.00003 s−1. In general, the true stress is affected by strain rate and temperature. The true stress declines with the increased temperature. The reason is that the thermal motion of atoms is enhanced, which decreases the bonding force of atoms, and plastic deformation easily occurs [The flow stress curves of the stretched alloy are displayed in y occurs ,32,33. Ty occurs ,35. Then−1. In −1 and 0.00003 s−1 when the true strains are 0.115 and 0.048, respectively. This reveals that the influence of dynamic softening is strong at lower strain rates or higher temperatures. Usually, the work hardening rate is used to depict work hardening behavior in deformed alloys. The plots of work hardening rate and true strain in diverse deformation conditions were acquired by differentiating the flow stress curves, as presented in θ < 15° and θ > 15° are referred to as LAGBs and high-angle grain boundaries (HAGBs), respectively. Meanwhile, the number of substructures decreases with the raised temperature. In addition, small fine grains around the elongated original coarse grains can be observed at 250 °C, which are marked with the white dotted line region in −1 to 0.00003 s−1. Additionally, some DRX grains can be found at 0.00003 s−1 in The inverse pole figure (IPF) maps of the stretched alloy at tested conditions are displayed in −1. The maximum mean misorientation angle is 8.40° at 250 °C. The volume fractions of HAGBs are 4.17%, 5.37%, 7.65%, and 17.10%, respectively, at 100, 150, 200, and 250 °C. In −1 to 0.00003 s−1, the mean misorientation angle increases from 5.08° to 6.71°, and the volume fraction of HAGBs increases from 6.69% to 12.41%. The distribution of grain boundary misorientation angles in different deformation conditions was evaluated by Channel 5 software (2009), as presented in According to −1 to 0.00003 s−1, the green regions decrease, and the value of KAM drops from 1.82° to 1.20°. The value of KAM is low at high temperatures or low strain rates, which reveals that the geometrically necessary dislocation density is low. This is because the DRV becomes more and more obvious, which accelerates the dislocation annihilation. Furthermore, the decreased substructures reduce the flow stress.The kernel average misorientation (KAM) maps in the tested conditions are displayed in 1 resulted from the dissolution of G.P. zones, while peak A2 was induced by the dissolution of θ′′ phases. Peak B is related to the precipitation of θ′ phases. Because of the annihilation and overlap of peaks, the dissolution peaks of Cu-rich clusters, as well as the precipitation peaks of G.P. zones and θ′′ phases, cannot be easily observed. The temperature ranges of the dissolution peak of Cu-rich clusters, the precipitation peak of G.P. zones, and the precipitation peak of θ′′ phases are 50–85 °C, 105–125 °C, and 175–190 °C, respectively. The four tested deformation temperatures  correspond to the four positions  marked in the curves. Cu-rich clusters and G.P. zones formed after the alloy were heated to 100 °C and 150 °C, respectively. θ′′ phases are presented in the matrix when the alloy was heated to 200 °C and 250 °C. Thus, the influences of heating process before deformation on the type of precipitates are significant. This is further discussed following the TEM observation.Al position. These are some main characteristics of G.P. zones formed during heating [Al position, revealing the precipitation of θ′′ phases [ heating ,48. When′ phases ,50.A1 and A2, which indicates the dissolution of G.P. zones and θ′′ phases. Peak B is correlated with the additional precipitation of θ′ particles. The decreased strain rate increases the characteristic temperature of peak A. This is because the migration of vacancies is more sufficient, and more G.P. zones transform to θ′′ phases. Thus, the stability of precipitates is increased when the strain rate is decreased. Therefore, peak A moves toward a high-temperature region, while the characteristic temperature of peak B moves toward a low-temperature region. This is because the nucleation of θ′ phases is affected by dislocations, and the deformation energy storage is low at low strain rates. The area of peak B decreases due to the decreased untransformed θ′′ phases. However, when the strain rate is 0.00003 s−1, it is hard to distinguish the endothermic peak and exothermic peak B from DSC curves at 200 °C and 250 °C . The experimental results are shown in The area of peak B in the tested deformation conditions was calculated. The RVF of θ′ phases can be evaluated as follows :(1)f=A1−−1, the type, content, and size of precipitates are affected by the deformation temperature, as shown in Al position in the SAED pattern reveal the appearance of G.P. zones. In Al direction due to the effects of tensile force. This is the so-called stress orientation effect [Al site indicate the formation of θ′ phases [Al direction. Except for the deformation temperature, −1 to 0.00003 s−1. The diffraction spots show that the precipitates are mainly θ′ phases. In addition, the distribution of these precipitates is not uniform, but most θ′ phases precipitate along the {020}Al direction.n effect ,52. The ′ phases ,46,53. θAl direction due to the stress orientation effect, which results in the uneven distribution of phases.Therefore, the type, content, and size of precipitates are affected by strain rate and deformation temperature. According to the above analysis, in the heating process before tensile deformation, the transformation of precipitates (G.P. zones→θ′′ phases→θ′ phases) is more sufficient at higher temperatures. During tensile deformation, the content and size of precipitates further increase due to the enhanced activation energy at lower strain rates or higher deformation temperatures. Meanwhile, the transformation rate of precipitates is accelerated, resulting in the increased θ′ and θ′′ phases. However, θ′ and θ′′ phases prefer to precipitate along the {020}−1. As the temperature is increased from 200 °C to 250 °C, the number of dimples decreases, and the inhomogeneity of dimple sizes increases. It can be concluded that the plastic deformation is more uneven with the increased temperature.The flow stress monotonically rises at 100 °C and 150 °C due to the dominant work hardening. When the temperature is raised to 200 °C and 250 °C, the enhanced dynamic recovery reduces substructures. The dynamic recovery becomes more obvious at lower strain rates. Meanwhile, some dynamic recrystallization grains appear due to the rotation of small substructures. Thus, the true stress firstly increases and then keeps stable or drops with increasing true strain.The type of precipitates is greatly affected by the heating process before deformation. When the alloy is heated to 100 °C and 150 °C, Cu-rich clusters and G.P. zones appear. When the alloy is heated to 200 °C and 250 °C, θ′′ phases precipitate.Al direction due to the stress orientation effect, which results in the uneven distribution of phases.The content and size of precipitates increase due to the enhanced activation energy at lower strain rates or higher deformation temperatures. Meanwhile, the transformation of precipitates is accelerated, and θ′ and θ′′ phases increase. However, θ′ and θ′′ phases prefer to precipitate along the {020}−1 and 150 °C, respectively.To alleviate work hardening and avoid the inhomogeneous precipitation of phases, the reasonable strain rate and temperature are about 0.0003 sThe deformation behavior and precipitation features in a stretched 2219 aluminum alloy at intermediate temperatures were studied. Several important conclusions are given below.


Magnaporthe oryzae (syn. Pyricularia oryzae) expressing the AVR-PikD effector, functions via receptor cooperation, with effector-triggered activation requiring both NLRs to trigger the immune response. To investigate the mechanism of Pikp-1/Pikp-2 activation, we expressed truncated variants of these proteins, and made mutations in previously identified NLR sequence motifs. We found that any domain truncation, in either Pikp-1 or Pikp-2, prevented cell death in the presence of AVR-PikD, revealing that all domains are required for activity. Further, expression of individual Pikp-1 or Pikp-2 domains did not result in cell death. Mutations in the conserved P-loop and MHD sequence motifs in both Pikp-1 and Pikp-2 prevented cell death activation, demonstrating that these motifs are required for the function of the two partner NLRs. Finally, we showed that Pikp-1 and Pikp-2 associate to form homo- and hetero-complexes in planta in the absence of AVR-PikD; on co-expression the effector binds to Pikp-1 generating a tri-partite complex. Taken together, we provide evidence that Pikp-1 and Pikp-2 form a fine-tuned system that is activated by AVR-PikD via receptor cooperation rather than negative regulation.Plant NLR immune receptors are multidomain proteins that can function as specialized sensor/helper pairs. Paired NLR immune receptors are generally thought to function via negative regulation, where one NLR represses the activity of the second and detection of pathogen effectors relieves this repression to initiate immunity. However, whether this mechanism is common to all NLR pairs is not known. Here, we show that the rice NLR pair Pikp-1/Pikp-2, which confers resistance to strains of the blast pathogen  Like animals, plants are constantly threatened by pathogens and pests. To defend themselves, they have evolved a sophisticated immune system that relies on both cell surface and intracellular receptors , 2. The R), establishing the TIR-NLR, CC-NLR and CCR-NLR families), a central NB-ARC domain , and a C-terminal leucine-rich repeat (LRR) domain. Conceptual frameworks for the roles of each domain are established, although their precise role may vary from one NLR to another [The canonical architecture of plant NLRs consists of an N-terminal Toll/Interleukin-1 receptor homology (TIR) domain or coiled-coil (CC) domain  and m another , 11). Th another –15. With another , 17. Mut another –20. Muta another . Mutatio another –26). The another –29, a fu another –32, but  another .NLRs can function as singletons, capable of both perceiving effectors and executing a response , 35. ThiArabidopsis are well established models in the study of genetically linked NLR pairs [CC-NLRs RGA5 and RGA4 from rice, and TIR-NLRs RRS1 and RPS4 from LR pairs , 48, 55.LR pairs , 56.Magnaporthe oryzae (syn. Pyricularia oryzae) carrying the AVR-Pik effector. Similar to RGA5, Pik-1 has an integrated HMA domain, but unlike RGA5 this is positioned between the CC and NB-ARC domain, rather than after the LRR. The Pik-1 integrated HMA domain directly binds the AVR-Pik effector [In rice, the CC-NLR pair Pik-1 and Pik-2 confers resistance to effector . HoweverNicotiana benthamiana [N. benthamiana experimental system. We show that intact, full-length, Pikp-1 and Pikp-2 are necessary for a cell death response upon effector perception. Truncation of any domain results in lack of effector-dependent cell death compared to wild-type. Further, expression of any specific NLR domain, or combination of domains, does not result in cell death. We also show that native P-loop and MHD-like motifs are required in both Pikp-1 and Pikp-2 proteins for receptor activity. Finally, we demonstrate that Pikp-1 and Pikp-2 are able to form homo- and hetero-complexes in planta in the absence of the AVR-PikD. Upon binding of the AVR-PikD effector, a tri-partite complex is formed that may represent the activated state of the receptor.We previously showed that the AVR-Pik elicited hypersensitive cell death mediated by the Pik NLR pair can be recapitulated using transient expression in leaves of the model plant thamiana , 57, 58.mas promoter and with C-terminal epitope tags  using the Golden Gate system [H46E constructs used were as described in [Agrobacterium tumefaciens strain GV3101 via electroporation.Domesticated sequences of full-length Pikp-1 and Pikp-2 (as described in ), and MLe system . To obtaribed in . All DNATo generate Pikp mutants (P-loop and MHD-like motifs), we introduced mutations into the relevant NB-ARC domain modules using site-directed mutagenesis. Subsequently these domain constructs were used to generate full length NLRs by assembly using the Golden Gate system. Each of the constructs were assembled with the CaMV35S promoter with relevant tags (6xHis + 3xFlag (HellFire (HF)) for Pikp-1 derivatives and 6xHA tag for Pikp-2 derivatives).Agrobacterium tumefaciens strain GV3101 carrying the appropriate constructs were suspended in infiltration buffer  and mixed prior to infiltration at the following final OD600: NLRs and NLR-derivatives 0.4, effectors 0.6, P19 (silencing suppressor) 0.1. Bacteria were infiltrated into leaves of ~4 weeks old N. benthamiana plants using a 1ml needleless syringe. At 5 days post infiltration (dpi), detached leaves were imaged under UV light on the abaxial side, and visually scored for cell death response (see below). To confirm protein expression, representative infiltration spots were prepared, frozen in liquid nitrogen, ground to a fine powder, mixed with extraction buffer (see below) in 2 ml/g ratio, centrifuged, mixed with loading dye and loaded on an SDS-PAGE gel for western blot analysis.https://www.r-project.org/) and the graphic package ggplot2 [Pictures of the leaves at 5 dpi were taken as described previously  and cell ggplot2 . Dots reProtein extraction was conducted as described in  with minWestern blots were performed as described previously . FollowiN. benthamiana using Agrobacterium tumefaciens mediated transformation (henceforth agroinfiltration). All constructs were tagged at their C-terminus with the HellFire tag (6xHis + 3xFlag (HF), for Pikp-1 domains) or HA tag (for Pikp-2 domains). The boundaries of the domains used were as defined in [To investigate the roles and requirements for individual domains of Pikp-1  and Pikp-2  in triggering cell death, we transiently expressed each of these in fined in .We found that each of the individual domains of either Pikp-1 or Pikp-2 were unable to trigger cell death when expressed alone, or in the presence of the corresponding paired NLR and/or effector . We confK296R and Pikp-2K217R). Such mutations restrict nucleotide binding, and have previously been shown to impair NLR function [N. benthamiana via agroinfiltration, we found that these mutations abolish cell death activity in planta, including when expressed in the presence of the paired NLR and the AVR-PikD effector  and a Pikp-2D559V mutant. On expression in N. benthamiana via agroinfiltration, we found that expression of these mutants alone did not result in auto-activity and cell death  served as a negative control for interactions. Each FLAG-tagged protein was expressed, and immunoprecipitated as expected  does not affect the homo-association of Pikp-1 or Pikp-2 , an inty linked , 77. Piky linked . So why Residues of the MHD-like motif are involved in binding ADP in the inactive state of NLRs , 78, andPlant NLRs can form both homo- and hetero-complexes both prior to and after effector recognition , 79, 80,In summary, our findings reveal that the Pikp-1/Pikp-2 NLR pair function via receptor cooperation rather than a suppression/activation mechanism, and signalling in planta requires the full-length proteins with native sequences at the P-loop and MHD-like sequence motifs. This suggests multiple mechanisms of regulation exist for NLRs. It is important to further investigate these mechanisms if we are to fully understand NLR function and use this to engineer improved disease resistance phenotypes in crops.S1 File(DOCX)Click here for additional data file.S1 Raw images(PDF)Click here for additional data file.


The objectives of the present investigation are to study the interaction and optimize the blend composition of flour of grinded Chia seeds, combined to Soy lecithin, a bread making improver, in a way to enhance the nutritional/functional value of bread without impairing its technological quality and to delay its staling rate. Nine formulations were prepared following a Central Composite Design. Technological attributes were evaluated both for fresh and stored bread. In the Response Surface Methodology (RSM) a desirability function identified the optimum doses of chia and lecithin incorporation to obtain the highest specific volume and the lowest crumb firmness. Compared to the control, samples with chia and lecithin significantly increased the nutritional value of bread. An innovative and interesting synergy was found in lecithin/chia combination to enhance the specific volume, to reduce the initial crumb firmness and to delay bread staling by retarding crumb firmness and reducing its water loss during storage. Using the RSM, the optimum blend containing (4.04%-Chia/1%-Lecithin) showed fresh bread with maximum specific volume and minimum crumb firmness. Whereas, bread combining the optimum blend (3.43%-Chia/1%-Lecithin) and stored for two days at room temperature showed the minimum crumb firmness.  Worldwide, consumption of soft wheat makes it a primordial cereal which occupies an important position in the international nutrition. In raw state, this cereal is the source of a large amount of nutrients required by the human organism. However, processing, which is prerequisite for manufacturing attractive products and obtaining a food in a suitable form and good palatability, may decrease the levels of the bioactive components in grains and also alter the bioavailability of these components . This isStarch retrogradation, one of the major factors of staling, includes the short-term development of amylose gel network structure  and a long-term reordering of amylopectin, which is a much slower process involving the outer branches recrystallization of this polymer ,4. StaliSalvia hispanica L). Chia is a summer annual plant belonging to the Labiatae family [Today, several anti-staling agents used in the bread making industry provide the opportunity to delay its quality properties degradation. Soy lecithin is an anti-staling agent which has an influence on starch retrogradation which is the key factor contributing to staling. This emulsifier acts by the formation of inclusion complexes with amylose or the outer branches of the amylopectin molecule, and delaying the swelling of starch granules . On the e family . Now thae family .Salvia hispanica L) combined with soy lecithin to wheat flour bread, second, to optimize the blend composition in a way to enhance the nutritional value of wheat flour bread, without impairing its technological quality.Due to the fact that the quality of a bread product mainly lies in the content and quality of the proteins that make up gluten, the substitution of wheat flour with other ingredients, could alter its cohesive and viscoelastic properties during the kneading. Consequently, the resulting dough loses its ability to retain gas during fermentation, meaning the bread produced is denser, with lower volume and a more compact crumb structure . These r−4 J. Chia seeds  were kindly granted by Primaria Premium Raw Materials S.L. . Soy lecithin was provided by SIPA . Compressed yeast  was used as a starter for the bread-making process.Commercial Spanish wheat flour , was purchased from the local market. The flour alveograph parameters were tenacity, P: 60 ± 1.5 mm; extensibility, L: 62.6 ± 2.8 mm; and deformation work, W: 148.6 ± 2.3 102 central composite rotational design (CCRD) of a Response Surface Methodology (RSM). The quantities added ranged from 0 to 10% on flour basis of chia and from 0 to 1% on flour basis of Soy lecithin. Nine assays were conducted including four factorial points (22) and five repetitions of the central point (Chia seeds flour and soy lecithin amounts (the two independent variables) were added to the formulation according to a 2al point . The stuProximal composition was determined in triplicate for fresh bread derived from each formulation. Protein determination was carried out by the Kjeldahl technique. Lipid content was extracted with petroleum ether under reflux conditions by the Soxhlet technique , whereasTo evaluate the impact of the incorporation of chia and lecithin on the mixing properties for each formulation, a Farinograph  was used, following the official standard method with slight modifications .The bread control dough formula consisted of: wheat flour (500 g), compressed yeast (2.5% flour basis), sodium salt (1.6% flour basis), and distiller water . The ingredients were mixed for 4.0 min, rested for 10 min, divided (100 g), kneaded and then rested (15 min). Doughs were manually sheeted and rolled, proofed during 1 h (at 28 °C and 85% relative humidity) and baked at 220 °C/25 min. The chia seeds were added at 5% and 10% on the flour basis and the soy lecithin at 0.5 and 1.0% on flour basis according to the formulations described in Bread quality was assessed for fresh and stored bread. Bread loaves were stored in polyethylene bags at 25 °C for four days. All the experiments of this part were done in triplicate.Bread volume was determined by a rapeseed displacement method and the average specific volume (volume/weight) was calculated. Moisture content evaluation was carried out both for the whole bread loaves and the bread crumbs, according to the American Association of Cereal Chemists (AACC) Method .Texture profile analysis was carried out using the Texture Analyzer . Two 25-mm thick slices were compressed in the center of the Texture Analyzer platform using a cylindrical probe of 36 mm in diameter under the following conditions: speed of 1.7 mm/s for the test; speed of 10 mm/s for the post-test; 40% compression and 5 g trigger force. Firmness, springiness, cohesiveness and chewiness were recorded . p < 0.05). In every case, the quadratic model was the most adequate accurate model. Specific volume and crumb firmness are the most critical attributes of bread. RSM was used to optimize the composition of chia and lecithin blends. A numerical desirability function was used to find the optimal composition in order to obtain bread with maximum specific volume and minimum crumb firmness values. Experimental data were fitted to the quadratic model:0 is a constant; b1 and b2 express linear effects of each variable; b3 and b4 are square coefficients; and b5 shows the interaction between the variables. Response surfaces of the models were plotted as a function of the two variables.Different models were studied to fit the experimental data using a MiniTab 17 software. The analysis of variance (ANOVA) was used to evaluate the significance of each equation , daily consumption of dietary fiber should be between 30–40 g. Its presence in the human diet helps to accelerate intestinal peristalsis, reduces the absorption of cholesterol and triglycerides, decreases glucose levels in the blood, and reduces the hunger sensation ,22. In t3/g. Compared to control, bread produced by the exclusive addition of lecithin at 1% presented a slight increase of the specific volume. This result may be explained by lecithin dough strengthening effect, which is not fully understood [Fresh bread specific volume was determined one hour after taking it out from the oven. The different loaves specific volume ranged from 3.22−3.45 cmderstood ,24.One theory suggests that lecithin potential to confer strength to wheat dough is due to the creation of a lecithin-gluten complex . This emBread containing only chia flour at 10% showed a specific volume similar to the control sample despite gluten dilution. In contrast, Coelho & Salas-Mellado  showed t3/g. The most important result to note is the positive effect of chia and lecithin combination on the specific volume. Regardless the added amounts, for the two formulations (Ch5-Lec0.5and Ch10-Lec1), the specific volume increased nearly to the same value 3.45 and 3.44 cmTherefore, we can conclude that lecithin incorporation not only corrected gluten proteins weakness caused by chia flour substitution, but also improved and strengthened these proteins . A higher moisture content leads to a better quality of the crumb during storage, meaning a softer bread crumb. The moisture decrease affects the crumb firming rate since cross links will be created between partially solubilized starch and gluten proteins .In the current study, moisture content in fresh bread crumbs decreased slightly with the following formulations Ch10-Lec0 > Ch0-Lec1 > Ch5-Lec0.5, in comparison with control fresh bread (43.7%) . Although the important ability of chia flour to absorb water during dough mixing, fresh bread crumbs presented lower moisture. This may be explained by the fact that the integrated chia flour absorbs water during dough formation and easily releases it during cooking . HoweverThus, the importance of chia mucilage water retention capacity appears during storage. The property of decreasing water loss in this phase of the process was also described by some studies. Likewise, Vázquez-Ovando et al.  reportedFrom another side, compared to reference bread, only fresh breads of the formulation Ch10-Lec1 presented higher crumb moisture (44.3%) which maintained its level until the end of storage (41.17%). Thus, only the interaction of lecithin at 1% and chia at 10% resulted in a stronger water holding capacity during baking and storage. This result may be explained by a cooperative effect between the two components. Lecithin role began from the mixing step by the adsorption onto the starch surface which might not allow the starch granules to take up water released by gluten after baking to the same extent as the control bread . TherefoBreads were daily submitted to texture analysis. Since consumers perceive bread with a softer crumb as more fresh, crumb firmness is one of the most critical texture parameters. At day 0, except bread sample containing only chia flour at 10%, which presented the highest crumb firmness (10.81 N), all the other bread samples baked either by a combination of chia and lecithin or by the exclusive incorporation of lecithin had a significantly lower firmness (between 4.36 and 5.6 N), as compared to reference bread crumb (7.47 N) . These rOn one hand, increasing crumb firmness by chia incorporation is mainly due to features related to dough gluten formation. Added flour dilutes the gluten matrix and causes crumb cell structure weakening. In fact, bread is a soft solid formed by two phases, a fluid one which corresponds to the air and a solid phase that corresponds to the gas-cell wall material. The solid phase is fully interconnected and the nature of the connectivity determines bread mechanical properties ,4. ConseOn the other hand, Soy lecithin addition resulted in lower crumb firmness which can be explained by its capacity to improve air entrapment. A similar effect has also been proven for other bread emulsifiers such as the diacetyl tartaric acid esters of monoglycerides (DATEM) and the sodium stearoyl-2-lactylate (SSL) . The intDuring storage, all bread crumbs’ firmness increased obviously as a result of staling. However, the same initial tendency was observed for all of them and the samples with improvers (except bread baked with 10% chia) maintained less firm crumbs. Also, it is interesting to note that, for Ch10-Lec1 sample, firmness increment was clear between day 0 and day 1, but henceforth crumb firmness increases in a non-significant way. At day 4, the highest values went to the control (30.95 N) and Ch10-Lec0 sample (32.79 N), with no statistical differences. Thus, despite gluten dilution at 10% of chia, final firmness of the correspondent bread crumb is statistically the same as the control bread. However, the sample with the exclusive addition of lecithin Ch0-Lec1 showed lower crumb firmness value (22.24 N), as compared to the reference. This value further decreased to reach 16.9 N in the sample containing chia and lecithin at maximal doses Ch10-Lec1 .All these observations confirm the anti-staling effect of lecithin and the combination of lecithin and chia. Initially lecithin incorporation produced lower crumb hardness and a retarded the staling rate through its interaction with amylose, as well as with amylopectin. Lecithin—with a higher content of lysophospholipids—inhibits crumb firming, associated with staling by complexing with starch amylose .The interactions between the lecithin and starch—adsorption process/complex formations— prevents starch from taking up water of the gluten network during the staling process . LecithiFrom another side, knowing chia flour mucilage high water retention capacity, chia incorporation in the formulation Ch10-Lec0 was suspected to decrease water loss during storage and thus to retard staling. However, the contrasting results permit the conclusion that the diluting effect of chia flour on gluten network dominate water retention softening effect. This confirms the fact that lecithin-chia combination in the two formulations Ch10-Lec1 and Ch5-Lec0.5 allowed to correct the firming effect of chia. In summary, amphiphilic character of lecithin providing the possibility of forming complexes with starch, proteins, shortening and water , coupledConcerning the other textural attributes, except bread made with 10% chia for which crumb chewiness was the highest (6.46 N), regardless of the added doses, the study showed that the bread either with chia and lecithin or with exclusive lecithin addition requires less work during fragmentation (from 3.37 N to 4.18 N) as compared to the control (6.03 N). This decrease of crumb chewiness is favorably perceived by consumers.Cohesiveness of all enriched bread crumbs increased significantly up to 0.78. As reported by Onyango et al. , cohesivIn general, bread textural properties  are unfavorable for consumption at day 2 onward. However, our results confirm that fresh bread of Ch10-Lec1 formulation had the lowest crumb firmness which was maintained until the end of storage with no significant difference between day 2 and day 4. In contrast, control crumb firmness varied between 15.76 N on day 2 and 30.95 N on day 4.To better understand and evaluate the interaction between crumb moisture content and its firmness, a cubic polynomial model was fitted to the experimental values. The adjustment trend confirmed the negative relation between the two variables. The more the crumb moisture increases, the more its firmness tends to decrease. This result was expected, seeing our results of moisture and firmness evolution mentioned above and knowing the plasticizing effect of water .The obtained responses  were statistically analyzed using the RSM, verifying the possibility of confirming the effects of chia and lecithin addition, mentioned previously, through mathematical models. Regression coefficients and variance analysis of the quadratic regression fitting models showing the relationships among responses and independent variables (chia and lecithin amounts) are presented in p < 0.01). This means that the effect of lecithin-chia interaction on reducing crumb firmness is more pronounced during storage than for fresh bread. Examining the interaction coefficients, it can be confirmed that the mutual influence of chia and lecithin to increase the specific volume of bread was significant. However, crumb firmness (day 0 and day 2) was negatively affected by this interaction, which was more significant at day 2 of storage   and maximum specific volume (3.45 cm3/g) . Otherwise, if the aim is to retard bread staling and obtain bread with minimum crumb firmness (10.24 N) after two days of storage, the optimal blend should contain  c. ExperiThe current study aimed to present that in addition to the functional and nutritional improvement of chia-added bread, an enhancement of fresh bread technological quality and a better resistance to staling during storage at 25 °C, was observed.In fact, incorporating chia flour at 10% decreased bread flour specific volume, whereas, bread produced by lecithin and Chia addition was characterized with an increased specific volume. Also, only chia and lecithin combination resulted in a stronger water holding capacity during baking. Besides, crumb firmness increased with chia addition and decreased with lecithin incorporation. However, there is a synergistic effect between the two additives to decrease crumb firmness. Another fact which should be considered is the good effect on crumb textural attributes. A significant enhancement of cohesiveness and a reduction of chewiness of all enriched bread crumbs were observed.During storage, regardless the added amounts, lecithin and chia combination resulted in a significant water loss and crumb firmness decrease, compared to samples containing only lecithin or only chia. This result confirms the synergy to increase water holding capacity and to delay bread staling.As a perspective, the optimum blends of chia and lecithin may be used to make bread at different heating rates in order to study the effect of baking conditions on bread staling and to optimize the temperature and the time of cooking. Also, our results may be tested in another baking technology. For example, it will be interesting to study the effect of the optimal composition on the staling rate of part-baked bread.


Mucocele of the appendix is the accumulation of mucoid material in the appendiceal lumen. Although the terminology is imprecise, as it does not differentiate between the benign and malignant nature of the condition, preoperative recognition is imperative as spillage of the mucus during surgical handling can result in grave complications like pseudomyxoma peritonei. Mucocele developing in a stump of the appendix, i.e., a remnant of appendiceal tissue after surgical removal of an inflamed organ, is an extremely uncommon phenomenon, as not many cases are reported in the literature. In this review, all cases reported in English literature are discussed. Mucocele developing into an appendiceal stump is an uncommon lesion, as only a few cases are reported. Recognition of this condition can be challenging because of its rarity and nonspecific symptoms –4. The sA thorough search of English literature was carried out with terms including stump, stump appendicitis, mucocele, and mucocele in stump in PubMed, Embase, and Google scholar and the references of all articles. More than 200 cases are reported in English literature for stump appendicitis after an initial appendectomy in the past and then developing into one of the complications like a mucocele in the stump. The data of all the articles were collected including history, examination, radiological findings, especially CT (computed tomography), operative details, and histopathology in a tabulated form.Ten cases of mucocele in the stump of the appendix were found in English literature. Tables Although first mentioned by Karl Freiherr von Rokitansky in 1842, Fere in 1877 coined the term mucocele and described it as appendicular dilatation secondary to accumulation of mucinous material in the lumen . FactorsThe incidence of appendiceal mucoceles has variously been reported as 0.07–0.3% of all the appendectomy specimen , 12, 18 Obstruction of the proximal end secondary to inflammation and confinement of mucus. Normal or ulcerated appendiceal mucosa lines the mucocele, and these are known as retention cysts , 21, simMucosal lining may change as seen in hyperplastic polyp of the colon .Sometimes, lining of the mucocele is composed of atypical (papillary epithelium) reminiscent of adenomatous polyp of the large bowel .Presence of cystadenocarcinoma where the epithelium is the same as in cystadenoma but demonstrates the features of stromal invasion by the neoplastic cells, and there may be cells in peritoneal deposits as mentioned by the Higa and others , 23, 24.Mucocele may result when an invasive adenocarcinoma (mucus secreting) affects the appendix , 19.Following mechanisms are proposed for the development of appendicular mucoceles and possibly stump mucoceles as well  , 19, 20.Hung mentioned obstruction of the appendiceal lumen by endometriosis and further exceptionally by inspissated mucus in cases of cystic fibrosis , 26. TheClinical manifestations are nonspecific; therefore, diagnosis is not usually straightforward. History of past appendectomy also poses difficulty and complexity in diagnosis. There is a reported delay of 4–30 years in diagnosis after the index appendectomy . PatientThere has been an association of the appendiceal mucoceles with increased incidence of colorectal neoplasms and to a lesser extent ovarian, endometrial, prostate, and other gastrointestinal malignancies, as reported by Fujiwara et al. . TherefoPreoperative localization along with characterization is important in decision-making and alarms the operating surgeon to be extremely careful in not spilling the contents in the peritoneal cavity. Spontaneous rupture has been reported leading to pseudomyxoma and signifies high chances of malignancy , 6. NotaIn the recent era, MDT (multidisciplinary teams) meetings are suggested to improve the management and outcome of all, particularly neoplastic diseases, so every case diagnosed preoperatively should be discussed, and appropriate management strategy is individualized. A surgical procedure in the form of limited ileocecal resection or right hemicolectomy is suggested as an optimum treatment for the mucocele of the stump appendix , 11, 25.Mucoceles in appendix are rarely encountered and in stump of appendix is further infrequent. A few cases are reported, and the most common presentation is with vague lower abdominal pain or slowly growing mass. US and CT scans are diagnostic in all cases, and surgical removal in the form of right hemicolectomy is the treatment of choice although stump appendectomy can be considered in smaller lesions. Resection should be carried out with utmost precaution to remove the mass intact as rupture may lead to catastrophic consequences. Multidisciplinary teams can play a vital role in managing the individual patients, and postoperative surveillance is needed as there are chances of other gastrointestinal neoplastic lesions in the future.