During reperfusion, transient ΔΨ depolarizations occurred concurrently with periods of increased mitochondrial oxidative stress . Supplementing the antioxidant system with GSH monoethyl ester suppressed ΔΨ oscillations  with 37% of mitochondrial clusters showing no ΔΨ oscillations . We found that limiting the production of reactive oxygen species using cyanide inhibited postischemic ΔΨ oscillations . Furthermore, ΔΨ oscillations were not associated with any discernable pattern in cell-wide oxidative stress or with the changes in cytosolic or mitochondrial Ca2+. Sustained ΔΨ depolarization followed cytosolic and mitochondrial Ca2+ increase and was associated with increased cell-wide oxidative stress. Collectively, these findings suggest that transient bouts of increased mitochondrial oxidative stress underlie postischemic ΔΨ oscillations, regardless of Ca2+ dynamics.Ischemia and reperfusion affect multiple elements of cardiomyocyte electrophysiology, especially within the mitochondria. We previously showed that in cardiac monolayers, upon reperfusion after coverslip-induced ischemia, mitochondrial inner membrane potential (ΔΨ) unstably oscillates between polarized and depolarized states, and ΔΨ instability corresponds with arrhythmias. Here, through confocal microscopy of compartment-specific molecular probes, we investigate the mechanisms underlying the postischemic ΔΨ oscillations, focusing on the role of Ca Reperfusion of the ischemic heart is crucial to limiting cell damage and initiating potential recovery, but it can also play a role in the injury process and trigger cardiac arrhythmias , 3, 4, 5via opening sarcolemmal ATP-sensitive K+ channels (sarcKATP) (ATP channels (with glibenclamide or glimepiride) does not prevent inexcitability during ischemia or the occurrence of reentry during reperfusion  regulation and consequently disrupting the cardiomyocytes’ electromechanical activity. Alternatively, the ionic imbalance, such as Ca2+ overload, might be behind ΔΨ oscillations or loss during reperfusion.A major organelle affected by IR is the mitochondrion, the main transformer of oxygen and nutrients into energy. In monolayer cultures of neonatal rat ventricular myocytes (NRVMs), we have shown that regional mitochondrial dysfunction alone, initiated by local perfusion with a chemical mitochondrial uncoupler, leads to inexcitability, slow conduction, and the occurrence of reentrant arrhythmias, mainly arcKATP) . FurtherarcKATP) , 9, we harcKATP)  depolariarcKATP) . The poserfusion . IR-induvia metabolic stress in the form of substrate deprivation  and decreased as mitochondria depolarized and the cells showed contracture . Occasionally, prolonged reperfusion led to sustained loss of ΔΨ, which was associated with further cytosolic GSH oxidation as the cardiomyocytes developed contracture (B). CytoGRX ratio increased to 50% of its maximum oxidized level 5.2 ± 1.03 min following 50% loss of cell-wide tetramethylrhodamine methyl ester (TMRM) signal (N = 10 experiments).During ischemia, cytoGRX F405/F488 ratio (GSSH:GSH) increased, indicating oxidation of cytosolic GSH pool, consistent with activation of the antioxidant pathway in response to accumulation of ROS during hypoxia. On reperfusion, an initial rapid increase in the oxidation of cytosolic GSH was observed, followed by partial recovery , A and Btracture B. CytoGRB). In contrast, measurement of the mitochondrial GSSG:GSH with mitoGRX revealed marked bursts of matrix GSH oxidation during individual ΔΨ oscillations (C–F). However, with more disorganized ΔΨ oscillations, GSH oxidation in the mitochondrial matrix and ΔΨ depolarization did not have a one-to-one relationship. An extended period of oxidative stress could be concurrent with several cycles of transient ΔΨ depolarization . These findings establish an interdependence between the level of mitochondrial oxidative stress and the occurrence of ΔΨ oscillations.There was no apparent connection between ΔΨ oscillations of individual mitochondrial clusters across the cardiomyocyte and the dynamics of cytosolic thiol oxidation. Close examination of the CytoGRX signal revealed no consistent discernable change in the cytosolic GSSG:GSH during ΔΨ oscillations B. In conllations , C–F. HoIn our recordings, the mitoGRX ratio increased concurrently with or shortly after ΔΨ depolarization. However, since the mitoGRX signal takes about 20 s to reach a plateau , ΔΨ depop = 0.027). In the presence of GSH-MEE, 37% of the analyzed mitochondrial clusters showed no ΔΨ oscillations during the first 15 min of reperfusion, compared to only 4% in control  significantly lowered the frequency of postischemic ΔΨ oscillations .We also tested whether cyanide could stabilize ΔΨ during reperfusion. Cyanide works by arresting electron flow at complex IV, resulting in the reduction of the FeS(III) electron acceptor of complex III. The lack of an oxidized acceptor shuts down the Q-cycle and inhibits ROS production at complex III , a major2+]m, followed by recovery in cardiomyocytes with a period of spontaneous contractions followed by quiescence  or reached a near-depolarization level after a few oscillations (D), while [Ca2+]m increased. ΔΨ fully depolarized upon the collapse of the whole mitochondrial network of the cardiomyocyte.These significant changes in i), we developed an adenovirus to express the FRET-based Ca2+ probe, Cameleon (CFP/YFP), in the cytosol (cytoCameleon). Mitochondria-targeted Cameleon (2+ levels ([Ca2+]m) during ischemia. To measure [Ca2+]m during reperfusion, we used the ratiometric genetically encoded probe . The cells were loaded with the potentiometric fluorescent dye, TMRM  for 1 h before the IR experiment when recording ΔΨ at high magnification using confocal microscopy . Dispersion of the TMRM signal throughout a cardiomyocyte was calculated and used to measure cell-wide ΔΨ, as described before  or mito-Grx1-roGFP2  mitoGRX on the sCameleon   on the center of the monolayer. This system restricts cellular access to oxygen and nutrients while metabolites accumulate in the limited extracellular space  the following: 135 NaCl, 5.4 KCl, 1.8 CaClar space , 11, 21.The changes in sarcolemmal membrane potential were mapped with a 464-photodiode array using the voltage-sensitive fluorescent dye, di-4-ANEPPS (5 μM), as the voltage waves, elicited by a stimulation pulse train (1 Hz), propagated through the monolayer. The data acquisition, processing, and visualization for the optical mapping setup were done using custom-made software written in LabVIEW .Monolayers were cultured on fibronectin-coated plastic coverslips (D = 21 mm) for 5 to 7 days. Using a previously described custom-built apparatus (https://imagej.net/software/fiji/)   was usedThe data are available from the corresponding author upon reasonable request.This article contains The authors declare that they have no conflicts of interest with the contents of this article.


Edema and ecchymosis after facial plastic surgery are a troublesome concern for both patients and surgeons. Corticosteroid administration is thought to shorten the recovery period and reduce these sequelae. Data regarding the efficacy of corticosteroid administration remains controversial among surgeons.We conducted this systematic review and meta-analysis to determine the effect of pre- and postoperative corticosteroids on postoperative complications in patients undergoing facial reconstructive surgery supported with different subgroup analysis.A comprehensive literature search of articles was conducted in PubMed, Cochrane Central, SCOPUS, and EBSCO through October 2019. We included all clinical trials in which patients underwent any type of facial plastic surgery to study the effect of corticosteroids on postoperative complications. We performed subgroup analysis according to the types and doses of corticosteroid preparation, in addition to a subgroup analysis of pre- or postoperative corticosteroid usage. All statistical analysis was performed using the RevMan software.p < 0.0001), but not for 8 mg corticosteroid (p = 0.06). Adding postoperative steroid dose to the preoperative one was associated with less edema and ecchymosis than preoperative administration alone.Nineteen studies were included in this systematic review, but only 10 of them were eligible for meta-analysis. The periorbital edema and ecchymosis scores were significantly reduced in the corticosteroids group compared to placebo −0.82, 95% CI , and -0.95, 95% CI , respectively. However, these significant differences were not maintained at day 3 and 7. Smaller doses of corticosteroid (8 mg and 10 mg) were associated with smaller differences in the mean score of upper and lower eyelid edema and ecchymosis, while the higher doses were associated with greater differences. Furthermore, preoperative corticosteroid usage significantly reduced the intraoperative bleeding when compared to placebo for higher doses > 50 mg per day (This comprehensive meta-analysis confirms a statistically significant benefit of preoperative corticosteroids. Furthermore, continuing the steroids postoperatively is associated with long-term reduction of complications. Higher doses of corticosteroids are associated with a more significant reduction in edema and ecchymosis, but further studies are recommended to determine the postoperative side effects, including surgical site infection and delayed healing. Facial plastic surgeries are one of the most commonly performed operative procedures worldwide. Facial surgery includes many types of procedures, including facelifts, rhinoplasty, and maxillofacial plastic surgery.The severity of edema and ecchymosis is the most important concern as it can delay the healing process of the involved tissues and alter the final intended aesthetic outcomeCorticosteroid administration is thought to shorten the recovery period and reduce postoperative edema and ecchymosis.9Data regarding the efficacy of corticosteroid administration remains controversial among surgeons, with a debate on the long-term benefit of intraoperative or postoperative corticosteroids administration in patients undergoing facial plastic surgery.Tuncel et al. 2013 reported that pre- and postoperative 10 mg dexamethasone with controlled hypotension considerably reduced postoperative swelling and ecchymosis following rhinoplasty, as well as intraoperative bleeding.Recently, in 2018, Sanober et al. showed that preoperative dexamethasone 8 mg could decrease periorbital edema by 50%, compared to placebo 33.3%. By the 7th day postoperatively, only 3.3% in the dexamethasone group had Grade III edema compared to 13.3%in the placebo group.Considering how common and important facial surgery procedures are, we conducted this systematic review and meta-analysis to resolve the conflicting conclusions reported earlier and to determine the effect of pre- and postoperative corticosteroids on postoperative complications in patients undergoing facial reconstructive surgery.We performed all steps of this systematic review in strict accordance with the Cochrane handbook of systematic reviews and meta-analysis.We searched the following medical electronic databases: PubMed, Cochrane Central, SCOPUS, and EBSCO through October 2019 using the following query: “”. We also searched the bibliography of eligible studies to find relevant articles.We included all prospective clinical trials  that met all the following criteria: a) Enrolled patients who underwent any type of facial plastic surgery; b) Studies that involved the use of corticosteroids compared or not with another intervention or placebo.We excluded: a) Observational studies; b) Studies with small sample size (less than ten patients); c) Studies performed on animal models; d) Reviews, case reports, conference abstracts, or case series; and e) Non-English articles and duplicate references.Eligibility screening was conducted in two steps, each by two independent reviewers : a) Title and abstract screening for matching the inclusion criteria, and b) Full-text screening for eligibility for meta-analysis. Disagreements were resolved employing the opinion of a third reviewer (Amani Obeid).Two independent reviewers  extracted the data and another reviewer (Jibril Yahya Hudise) resolved disagreements. The extracted data included the following: a) General characteristics of each study including; study setting, study design, sample size, type of intervention and the doses, type of competitor; b) Patients’ baseline characteristics of each study including: age, gender, race; c) Operation characteristics including: type of surgery, duration of surgery, follow-up period, and postoperative edema assessment; d) Outcomes of interest including swelling (edema), bruising (ecchymosis), intraoperative bleeding and e) Risk of bias criteria.To assess the risk of bias within the included clinical trial, two independent reviewers  used the Cochrane Risk of Bias (ROB) assessment tool for the randomized clinical trial, clearly described in (chapter 8.5) of the Cochrane handbook of systematic reviews of interventions 5.1.0.18The Cochrane risk of bias assessment tool includes the following domain: sequence generation (selection bias), allocation sequence concealment (selection bias), blinding of participants and personnel (performance bias), blinding of outcome assessment (detection bias), incomplete outcome data (attrition bias), selection outcome reporting (reporting bias) and other potential sources of bias. The authors’ judgment is categorized as “low risk”, “high risk” or “unclear risk” of bias.We calculated the Mean Difference (MD) and 95% Confidence Intervals (95% CI), if outcomes were measured in the same way between trials. Standardized Mean Difference (SMD) was calculated in the case of difference in the scoring system.In some trials, we extracted the data from graphs using PlotDigitizer software.Gurlek et al. 20062 less than 50%), the fixed-effect model applied to pool data. In the presence of statistical heterogeneity (I2 > 50%), the random-effects model for meta-analysis was applied.We tested for heterogeneity among included studies by the Chi-Square tests and quantified its extent by the I-Square test. In the absence of clinical and statistical heterogeneity  for Windows software.We performed subgroup analysis acceding to the types and doses of corticosteroid preparation, in addition to a subgroup analysis of per or postoperative corticosteroid.,Our search yielded a total of 1076 studies. Following screening and excluding duplicates, 89 studies remained that entered full-text screening. Finally, 19 studies16We reported an overall low risk of bias according to ROB assessment tool. A summary of risk of bias assessment was reported in We summarized the baseline characteristics of 19 included studies in Dexamethasone 8 mg was administered in five studies, while dexamethasone 10 mg was administered in seven studies. Lin et al. 2017Four studies investigated the methylprednisolone with different doses. Eleven studies investigated the efficacy of combined pre- and postoperative corticosteroid, while eight studies reported preoperative corticosteroid only. Fifteen studies reported the results after rhinoplastic surgery, two studies after orthognathic surgery,A total of 439 patients from 10 clinical trials were included in the meta-analysis. All of these studies reported the assessment of upper and lower eyelid ecchymosis and edema postoperatively using the four-point scale. The most commonly used corticosteroid doses were 8 mg and 10 mg. The higher doses were pooled in one group (corticosteroid > 50 mg). We pooled the results of postoperative ecchymosis and edema at day 1, 3, 5, 7 and 10. The detailed results of subgroup analysis according to the types and doses of the used corticosteroid as well as a subgroup analysis of pre and postoperative corticosteroids are presented in Supplementary Tables 1 and 2.p  = 0.33, I² = 10.6%. Pooled studies were homogenous for dose 8 mg (I2 = 0%), but heterogonous for dose > 50 mg (I2 = 69%). Heterogeneity was best resolved by exclusion of Tuncel et al. 2013 A (single dose of 10 mg/kg group), I² = 22% . Tests for subgroup differences between corticosteroids dose 8 mg, 10 mg, and > 50 mg were not significant, Chi² = 2.24, I² = 22% .Figure 3At day 3 postoperatively, corticosteroid was continuously reduced the score of upper eyelid edema compared to placebo  with no heterogeneity (I² = 0%), except for dose 8 mg subgroup (I² = 87%) . Test for subgroup differences between corticosteroids dose 8 mg, 10 mg, and > 50 mg was significant, Chi² = 6.73, 2 = 0%), but heterogonous for dose > 50 mg (I2 = 66%). Heterogeneity was best resolved by exclusion of Tuncel et al. 2013 A (single dose of 10 mg/kg group), I² = 6%  difference between corticosteroid (10 mg/kg) and placebo at day 5 postoperatively.In comparison to the placebo, corticosteroids reduced the mean score of upper eyelid ecchymosis significantly at day 1 postoperatively . Test for subgroup differences between corticosteroids dose 8 mg, 10 mg, and > 50 mg was not significant, Chi² = 0.39,  I² = 0% . The samp =  0.02, I² = 75.7% . Tests for subgroup differences between corticosteroids dose 8 mg, 10 mg and > 50 mg were significant, Chi² = 8.24,  = 75.7% . The samOn the first day, the overall effect size of the edema score revealed that edema was significantly reduced in corticosteroids group compared to placebo . This was not achieved in corticosteroid 8 mg group, but achieved in higher doses as presented in In comparison to the placebo, the corticosteroids group reduced the postoperative ecchymosis at day one  . While op =  0.47, I² = 0%)  without difference or heterogeneity between the corticosteroid doses  .Figure 9The present systematic review included 19 clinical trials. Of them, only 10 studies were included in the quantitative evidence synthesis. Our meta-analysis provides Class 1 evidence that corticosteroid has a beneficial effect in reducing postoperative edema and ecchymosis. In addition, preoperative corticosteroids decreased intraoperative bleeding. Although postoperative edema and ecchymosis have commonly occurred after facial plastic surgery in which steroids are commonly used, small numbers of studies were conducted to establish its effectiveness and determine the preferred dose and type to be administered.On the first postoperative day, smaller doses of corticosteroid (8 mg and 10 mg) were associated with smaller differences in the mean score of upper and lower eyelid edema and ecchymosis, while the higher doses were associated with greater differences. None of the included studies reported any adverse events related to the higher or lower doses of corticosteroid.p <  0.0001), but not for 8mg corticosteroid (p = 0.06). Adding postoperative steroid dose to the preoperative one was associated with a decrease in edema and ecchymosis than when given preoperatively alone. Using higher doses of methylprednisolone could improve the postoperative edema and ecchymosis efficiently at day one postoperatively rather than any other steroid. Using a higher methylprednisolone dose was not associated with any intraoperative or postoperative adverse events, but it significantly decreased the intraoperative bleeding.4Furthermore, preoperative corticosteroid administration significantly reduced the intraoperative bleeding when compared to placebo for higher doses > 50 mg per day . Grade 1 edema was present in the corticosteroid group at day 1 (43.2%), day 2 (29.5%), day 5 (13.6%) and was completely absent from 7th postoperative day onward. In the placebo group, on the 7th postoperative day, 33.3% of the patients still had Grade 1 and 16.7% had Grade 2 edema. Moreover, 16.7% in the placebo group continued to show Grade 1 edema on the 10th postoperative day.Alajami and colleaguesRegarding the intensity of ecchymosis and severity of edema, Totonchi et al. 2007Tuncel et al. 2013The overall quality of the evidence of the included studies is high. The main concern is the small sample size of each article. According to the American Society of Plastic Surgeons report 2018, Nose reshaping is one of 2018′s top 5 cosmetic surgical procedures .Although we are presenting the first large comprehensive systematic review (19 included studies) to investigate the effect of corticosteroids on reducing the post-facial plastic surgery complications, we reported a consistent result with the previous Cochran review in 2014Most of the included studies were heterogeneous regarding the numbers and dosage of pre-and postoperative corticosteroids as well as the type of the steroids used.. Studies by Abukawa et al.,In conclusion, this comprehensive meta-analysis confirms a statistically significant benefit of the preoperative administration of corticosteroid when compared to placebo. Furthermore, continuing the steroids postoperatively is associated with long-term reduction of complications, including edema, ecchymosis, and intraoperative bleeding. Finally, higher doses of corticosteroids are associated with a more significant reduction in edema and ecchymosis, but further studies are recommended to determine the postoperative side effects such as surgical site infection and delayed healing.The authors declare no conflicts of interest.


Here, we show that the ferritin levels are affected by RNase III and PNPase, which target the ftnA 3’UTR. Rifampicin mRNA stability experiments revealed that the half-life of the ftnA mRNA is affected by both RNase III and the ftnA 3’UTR. A transcriptional fusion of the ftnA 3’UTR to the gfp reporter gene decreased green fluorescent protein (GFP) expression, indicating that the ftnA 3’UTR could work as an independent module. Additionally, a chromosomal deletion of the ftnA 3’UTR impaired S. aureus growth under conditions of iron starvation. Overall, this work highlights the biological relevance of the ftnA 3’UTR for iron homeostasis in S. aureus.Iron acquisition and modulation of its intracellular concentration are critical for the development of all living organisms. So far, several proteins have been described to be involved in iron homeostasis. Among them, ferritins act as the major iron storage proteins, sequestering internalized iron and modulating its concentration inside bacterial cells. We previously described that the deletion of the 3’-untranslated region (3’UTR) of the  Staphylococcus aureus need to overcome the restricted availability of free iron, a process known as nutritional immunity , gene regulation, and DNA biosynthesis . When inimmunity . Staphylorldwide , has devorldwide .S. aureus, the ferric uptake regulator (Fur) and peroxide stress transcriptional regulator (PerR) are the two main transcriptional regulators that modulate the excess of iron and oxidative stress. When high levels of iron are sensed inside the cell, Fur inhibits the transcription of several genes involved in iron and heme uptake and promotes the expression of efflux pumps  that cause oxidative stress and damage DNA as well as other essential molecules . For thiux pumps . Alterna by PerR .icaR mRNA modulates the production of IcaR, the main repressor of PIA-PNAG exopolysaccharide biosynthesis in S. aureus. This 3’UTR contains a UCCCC motif that binds to the Shine-Dalgarno (SD) sequence at the 5’UTR of the same mRNA and inhibits ribosome binding and translation. This interaction results in the formation of a double-stranded RNA (dsRNA) substrate for endoribonuclease III (RNase III) to process  or by an mRNA processing site (type II) located at the 3’UTR (3’-untranslated regions (3’UTRs) have recently emerged as impactful post-transcriptional regulatory elements that regulate the levels of the mRNAs in which they are encoded through different mechanisms . For exa process . The 3’Ucleavage . Anotherg others . 3’UTRs radosome . The Salpression . Additiohe 3’UTR .S. aureus ftnA gene expression was modulated by its own 3’UTR. Deletion of the ftnA 3’UTR increased ferritin expression, portraying said 3’UTR as a putative post-transcriptional regulatory element  supplemented with 0.25% glucose (TSBg) or, when indicated, in modified chemically defined medium , and SuperBroth  media were used, respectively. For selective growth, media were supplemented with the appropriate antibiotics at the following concentrations: ampicillin (Amp), 100 μg ml−1 for E. coli transformants; erythromycin (Erm), 1,5 μg ml−1 or 10 μg ml−1 for S. aureus cells harboring the pMAD or pCN plasmids, respectively.Bacterial strains, plasmids, and oligonucleotides used in this study are listed in d medium . To prepMost of the plasmids used in this study were engineered as previously described . pMAD plListeria monocytogenes pAD-cGFP plasmid as a template  reporter plasmids were constructed using the template . To buil-GFP-rvs  and the -ftn-rvs  and inseterm-ftn . The amp3XFFtnAΔ3’UTR19-56 and 3XFFtnAΔ3’UTR57-93 were constructed using the p3xFFtnA plasmid backbone. For the 3XFFtnAΔ3’UTR19-56 plasmid, an overlapping PCR was performed using oligonucleotide pairs +1-ftn and 3’UTR-ftn-19-56-fw and 3’UTR-ftn-19-56-rv and term-ftn. Analogously, oligonucleotides +1-ftn and 3UTR-ftn-term-1/2 were used for the 3XFFtnAΔ3’UTR57-93 plasmid (The plasmids expressing  plasmid . The res plasmid .600). Erlermeyer flasks containing fresh TSBg+Erm were inoculated with precultures to reach starting OD600 of 0.02. For rifampicin mRNA stability assays, test cultures were grown at 37°C and 200 rpm until an OD600 of 0.5 . Six aliquots of 20 ml of the culture were transferred to 50 ml Falcon tubes containing 300 μg ml−1 of rifampicin and incubated at 37°C for 0, 2, 4, 8, 15, and 30 min. Then, 5 ml of stop solution (95% ethanol and 5% phenol) were added to the samples and centrifuged for 2 min at 4,400 g. Pellets were frozen in liquid nitrogen and stored at −80°C. RNA extraction and Northern blot analysis were performed as described in ftnA or the 3XFftnA mRNAs. The mRNA levels were quantified by densitometry of Northern blot autoradiographies using ImageJ.Precultures were grown in 5 ml of TSBg supplemented with Erm (TSBg+Erm) and incubated overnight (ON) at 37°C and 200 rpm. Bacterial concentrations of the precultures were estimated by measuring their optical density  indicate p-values lower than 0.05 (p < 0.05) while ns indicate not significant differences.Bacteria were grown as described above. At ODwo/Fe) overnight at 37°C and 200 rpm . OD650 measurements were performed every 30 min at 37°C for a period of 20 h.Precultures were grown in 5 ml of modified chemically defined medium without iron  when compared to the p3xFFtnA strain  from the WT and Δ3’UTR ftnA mRNAs, respectively. The selected RNase mutants included ∆rnc , ∆pnpA  of the ftnA mRNA downstream of the gfp gene  and 68–89 (conserved region II), respectively, besides the putative TT , or 57 and 93 (p3xFFtnA∆3’UTR57-93), which selectively eliminated conserved regions I and II, respectively . Northern blot results showed that the chromosomal ftnA∆3’UTR mutant expressed higher ftnA mRNA levels than the WT strain as it occurred with the plasmidic mutant  at 37°C and measured bacterial growth by registering the optical density every 30 min. However, no growth differences were observed between the WT and ∆3’UTR mutant in the MMwo/Fe , a strong iron chelator. ftnA∆3’UTR mutant, while adding 500 μM of DIP completely impaired it. Altogether, these data portrayed the ftnA 3’UTR as an essential module to control the ftnA mRNA expression and maintain proper iron levels for S. aureus growth under iron starvation conditions.Then, we aimed at comparing the capacities of the  MMwo/Fe . Since i MMwo/Fe . TherefoftnA 3’UTR, which is mainly targeted by RNase III and PNPase . Whether other sRNAs and/or asRNAs interact with the ftnA 3’UTR remains to be explored. Moreover, knowing that global regulatory RNA chaperones have been already shown to bind 3’UTRs  despite the ftnA CDS being conserved ; and the Spanish Ministry of Science and Innovation (PID2019-105216GB-I00) grants.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.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


There has been a growing interest in quantitative predictions of the intermolecular binding energy of large complexes. One of the most important quantum chemical techniques capable of such predictions is the domain-based local pair natural orbital (DLPNO) scheme for the coupled cluster theory with singles, doubles, and iterative triples [CCSD(T)], whose results are extrapolated to the complete basis set (CBS) limit. Here, the DLPNO-based focal-point method is devised with the aim of obtaining CBS-extrapolated values that are very close to their canonical CCSD(T)/CBS counterparts, and thus may serve for routinely checking a performance of less expensive computational methods, for example, those based on the density-functional theory (DFT). The efficacy of this method is demonstrated for several sets of noncovalent complexes with varying amounts of the electrostatics, induction, and dispersion contributions to binding  calculations). It is shown that when applied to dimeric models of poly(3-hydroxybutyrate) chains in its two polymorphic forms, the DLPNO-CCSD(T) and DFT-SAPT computational schemes agree to within about 2 kJ/mol of an absolute value of the interaction energy. These computational schemes thus should be useful for a reliable description of factors leading to the enthalpic stabilization of extended systems. A quantitative description of intermolecular noncovalent interactions is the key factor in understanding properties of gaseous and condensed phases , molecul60 atoms ), reduce60 atoms . Current60 atoms ,23,24,2560 atoms ,27. The ferences ,29,30 anferences ). Neverteference ; using seference ; and exteference . These asets see . This prtion see . Specifilo (QMC)  approachlo (QMC) ,41,42,43First the benchmark rrection  was applThe respective portions of the total interaction energy are the Hartree–Fock component, e review  for disce review  , instead of using the iterative scheme that would provide results denoted as (T1) [1) calculations with respect to their DLPNO-CCSD(T0) counterparts, and by negligible differences between the two sets of results for dimers from the Set3x6 (see It should be noted that the LPNO-MP2  correlat as (T1) . This cht3x6 see .The eference ). The li/mol see .The interaction energies computed using the focal point method from Equation (2) were also checked against yet another set of results. Namely, the total DLPNO-CCSD(T) energies were calculated while employing the aTZ; aQZ; a5Z series of basis sets, and extrapolated to their CBS limit using the mixed Gaussian/exponential form from reference . The undalue see . Figure alue see . This reα minimizes (in the least-squares sense) differences between While the composite procedure from the previous part (Equation (2)) is clearly successful in reliably predicting the interaction energies, it also requires results obtained using the aQZ basis set, which would be impractical for very large systems. Hence, an attempt was made to devise some less costly computational protocol. It is stressed that a direct application of Equation (2) together with smaller basis sets  cannot be expected to lead to an accurate estimate of the CCSDTCBS . InsteadCBS term . A partint study  and refent study  that wernt study  were appα = −4.473 (rounding to four digits is performed on the basis of an estimated covariance that is not shown). The correlation energies were treated in an analogous way. Specifically, an optimal value of the coefficient β for the MP2 correlation energy contribution was obtained by minimizing differences between The fit gives β = 2.796. As for the post-MP2 correlation energy term, numerical tests revealed that yet another coefficient would be needed, which is denoted as γ and minimizes differences between This value is γ = 2.741 for the parameter An optimal value of this coefficient is found to be The procedure from utations , while Gutations  are showeference  are emplThe DLPNO-based methods from  quality . Moreove quality  and refe quality , develop quality , or for  quality ). It caneference , are inceference . The ordractions . The comractions . Neverth2, anisole∙∙∙anisole, anisole∙∙∙ammonia, 1-naphtol∙∙∙acetylene, HCl∙∙∙HCl, benzene∙∙∙water, 1-naphtol∙∙∙ammonia, and HCl∙∙∙water. The MP2/aTZ geometries, which are provided in ‘geometries.tar’ file in 2 , acetyleference ), HCN∙∙∙eference ), NCH∙∙∙eference ), HCN∙∙∙eference ), and 1-eference  with defThe dimeric models of the α and β polymorphs of PHB were prepared using coordinates from references ,37, respThe configurations 1–7 of the furan∙∙∙toluene dimer were taken from ‘ja9b00936_si_002.xyz’ file of supporting materials to reference . CoordinThe SAPT-DFT, canonical CCSD(T) and MP2 calculations were carried out in Molpro 2021.2 . The samThe MP2/a5Z energies for an application of Equation (1) were obtained in the resolution-of-the-identity integral approximation ,73 whileThe DLPNO-based computations were carried out in ORCA 5.0 . The und® Optimization Toolbox™ was used to perform the nonlinear fitting of models expressed by Equations (3)–(5). The values of ® Fortran Library; the respective data files are included in The Levenberg–Marquardt algorithm as implemented in ‘lsqcurvefit’ function of MATLAB


Ocimum basilicum L. is an aromatic plant rich in bioactive metabolites beneficial to human health. The agronomic biofortification of basil with Zn could provide a practical and sustainable solution to address Zn deficiency in humans. Our research appraised the effects of biofortification implemented through nutrient solutions of different Zn concentration  on the yield, physiological indices , quality, and Zn concentration of basil cultivars ‘Aroma 2’ and ‘Eleonora’ grown in a floating raft system. The ABTS, DPPH, and FRAP antioxidant activities were determined by UV-VIS spectrophotometry, the concentrations of phenolic acids by mass spectrometry using a Q Extractive Orbitrap LC-MS/MS, and tissue Zn concentration by inductively coupled plasma mass spectrometry. Although increasing the concentration of Zn in the nutrient solution significantly reduced the yield, this reduction was less evident in ‘Aroma 2’. However, regardless of cultivar, the use of the maximum dose of Zn (50 µM) increased the concentration of carotenoids, polyphenols, and antioxidant activity on average by 19.76, 14.57, and 33.72%, respectively, compared to the Control. The significant positive correlation between Zn in the nutrient solution and Zn in plant tissues underscores the suitability of basil for soilless biofortification programs. A diversified and well-balanced diet based on high nutritional quality foods is prerequisite to good health and, according to the World Health Organization (WHO), it is dependent on the development of sustainable agricultural systems . Insidioad hoc Zn concentrations would allow a standardized, fine control of leafy vegetable quality as already observed in Lactuca sativa L., Thlaspi caerulescens, and Brassica oleracea (2+ through ZIP transporters or chelated with low molecular weight compounds (phytosiderophores), a mechanism typical only of Poaceae  , and reduction in yield are observed (–1 dry weight are toxic (–1), lettuce (20-60 mg kg–1), broccoli , and leafy greens (up to 700 mg kg–1) (Ocimum basilicum L.). The scientific literature has not classified basil as either a hyperaccumulator or non-hyperaccumulator species. In any case, in light of the encouraging results obtained with Selenium and Iodine biofortification programs  . In geneobserved . Howeverre toxic , causingre toxic . Althougmg kg–1) , to our programs  we hypotOcimum basilicum L.) ‘Aroma 2’  and ‘Eleonora’  were sown at a density of 317 pt m–2 on April 13, 2021 in peat and vermiculite (1:2 v/v) in 54-hole polystyrene trays  and grown in a floating raft system (FRS) in individual plastic trays filled with 35 L of nutrient solution (NS). As indicated by 3-, 1.5 mM P, 1.75 mM S, 3.0 mM K, 4.5 mM Ca, 1.5 mM Mg, 1.0 mM NH4+, 15 MM Fe, 9 MM Mn, 1.6 MM Zn, 0.3 MM Cu, 20 MM B, and 0.3 MM Mo. The trial was carried out in a randomized design with three replicates in a factorial arrangement (2 × 5), with two basil cultivars (Aroma 2 and Eleonora) and four biofortification treatments plus control. The latter consisted of four doses of Zn  using ZnSO4 × 7H2O  as a Zn source in the nutrient solution. Each experimental unit consisted of 54 plants. Biofortified nutrient solutions were provided twenty days after planting . The biofortified treatment lasted for 23 days. Plants were grown under natural light conditions. During the growing cycle, temperature and relative air humidity were recorded continuously with an interval of 10 minutes with dedicated WatchDog A150 dataloggers  placed at canopy level. Specifically, the day/night average air temperature and relative humidity were 27/18°C and 50/70%, respectively.The experimental trial was conducted at the Department of Agriculture, Federico II University,  in an unheated greenhouse from May 3 to 26, 2021. Genovese basil seedlings (–1). Leaf area was quantified using ImageJ v1.52a software . The epigeal parts of each plant were dried in a ventilated oven at 70°C for 3 days to determine the dry biomass of the shoots and roots (g plant–1) and their percent ratio. The dry matter (%) of the shoots was calculated as follows:Before flowering (43 days after sowing), twenty plants per replicate were sampled for the determination of height (cm), number of leaves, and fresh biomass  for mineral concentration determination. A representative plant sample was collected for each experimental unit and stored in liquid nitrogen for further qualitative analysis.Leaf color assessment according to human vision was performed using the CIELab color space defined by the CIE , which separates greyscale (L*) information more clearly from color (a* and b*) information. The color coordinates were measured on twenty young fully expanded leaves per replicate using a Minolta CR-300 colorimeter . Chroma and hue angle were calculated from the equations reported by 2 (ACO2), the transpiration rate (E), and the stomatal conductance (gs). Relative humidity (RH) and CO2 concentration of the leaf gas exchange analyzer were set at ambient values, while photosynthetically active radiation (PAR) and airflow rate were constant at 2,000 Mmol m–2 s–1 and 500 mL s–1, respectively. Chlorophyll fluorescence (Fv/Fm) measurements were made using a fluorometer Fv/Fm meter . The SPAD index was assessed using a Minolta SPAD-502 chlorophyll meter . All physiological measurements were performed between 09:30 and 12:00 am on five fully expanded healthy young leaves for each replicate.Leaf gas exchange measurements were performed with a Li-6400 hand-held analyzer . The measured parameters of interest were the net assimilation of CO–1 fresh weight (fw).Chlorophyll a and b concentrations were determined by UV-Vis spectrophotometry  with an absorbance of 647 and 664, respectively, as described by β-carotene and lutein concentrations were quantified by high-performance liquid chromatography with diode array detection (HPLC-DAD) after extraction according to –1 dw.The –1 dry weight (dw)] was determined by an inductively coupled plasma mass spectrometer  after digestion with a mixture of HCl (37%) and HNO3 (65%) . An appropriate calibration curve was prepared using a standard solution with 1.0 to 100 Mg L–1 Zn concentrations.According to the method described by + , DPPH , and FRAP (ferric reduction antioxidant potency) were determined by UV-VIS spectrophotometry  according to the protocols described by –1 dw. Phenolic Concentration DeterminationThe antioxidant activities ABTS–1 dw.One hundred milligrams of freeze-dried basil were used to quantify and determine polyphenols using a UHPLC system  equipped with a thermo-stated column  and a quaternary pump . Mass spectrometry analysis was facilitated by a Q Exactive Orbitrap LC-MS/MS system . Phenolic compound sampling was performed according to the protocol detailed by p< 0.05 level. SPSS 20 software package  was used. Data represent mean ± standard error of 3 replicates (n = 3).Data were subjected to analysis of variance (ANOVA) and means for cultivar treatment (CV) were compared by the Student’s t-test, whereas the means for the biofortification treatment (Zn) and the two-way interaction (CV × Zn) were compared using the Tukey-Kramer HSD test at the Plant height, number of leaves, fresh biomass, total dry biomass, and dry matter content for the control treatment were higher for ‘Aroma 2’ than ‘Eleonora’ Table 1.Significant CV × Zn interactions were observed for all leaf colorimetric parameters analyzed , since ‘Aroma 2’ exhibited a higher ACO2 than ‘Eleonora’  of ‘Aroma 2’ was not affected, transpiration of ‘Eleonora’ was reduced on average by 19.9% for treatments exceeding 12.5 MM Zn in the nutrient solution. Zinc treatments decreased SPAD index of ‘Eleonora’ by 12.3% on average, while only the intermediate Zn treatments (25 and 37.5 MM) reduced the chlorophyll concentration of ‘Aroma 2’ by 4.3%, on average. Also. Zn treatments negatively affected the maximum quantum yield of photosystem II (Fv/Fm) of both cultivars, namely by 4.2% and 3.9% on average, for ‘Aroma 2’ and ‘Eleonora’, respectively. However, while ‘Eleonora’ Fv/Fm appeared to decrease proportionally with increasing Zn concentration, ‘Aroma 2’ Fv/Fm decreased up to 25MM concentration and showed no further change thereafter.Cultivar physiology under control conditions was similar for all the parameters examined, except for COleonora’ Table 3.β -carotene values were higher in ‘Aroma 2’ than ‘Eleonora’, although the latter had a higher total chlorophyll concentration  and shoots (19.9%) than the ‘Eleonora’ control plants. Increasing Zn concentration in the nutrient solution resulted in both cultivars showing a relative increase in root and shoot Zn concentration. Overall, supplementing the nutrient solution with Zn increased root Zn concentration in ‘Aroma 2’ by 13.4% and ‘Eleonora’ by 12.8%, when compared to the control. Notable was also the differentiation in Zn concentration by the shoots of the two cultivars compared to the control, as Zn accumulation in the shoots increased on average by 23.1% in ‘Aroma 2’ compared to 9.7% in ‘Eleonora’. This disproportional Zn accumulation potential in the shoots of the two cultivars was manifested as significant CV × Zn interaction. In fact, Zn addition to the nutrient solution had a greater effect on ‘Aroma 2’ plant shoot Zn levels compared to the control, which was overall approximately 138.8% higher than that of ‘Eleonora’  was altered by supplemental Zn in the nutrient solution Table 4.−1 dw and 4319 Mg g−1 dw for ‘Aroma 2’ and ‘Eleonora’ respectively, while rosmarinic acid was the second most abundant phenolic acid (523.8 Mg g−1 dw for ‘Aroma 2’ and 358.5 Mg g−1 dw for ‘Eleonora’). The ranking of the remaining phenolic acids was also similar in the two cultivars except for salvianic and caffeic acid. Salvianic and caffeic acids ranked 7th and 9th for ‘Aroma 2’, while it was the other way around for ‘Eleonora’. Under control conditions, ‘Aroma 2’ total phenolic acids concentration was higher by 12.8% than ‘Eleonora’, since half of the individual phenolic acids concentration  was higher in ‘Aroma 2’ than in ‘Eleonora’. No differences among the two cultivars were observed for the rest of the phenolic acids  in the control solution. Zinc additional quantity to the nutrient solution altered the individual phenolic acids concentration and the total phenolic concentration of both cultivars. An exception was recorded for ‘Eleonora’ salvianolic acid L concentration which was not affected by Zn treatments. The effect of Zinc, though, was subjected to significant CV × Zn interaction for all the individual phenolic acids and for the total phenolic acids concentration. In all the phenolic profile parameters examined, interaction was significant because Zn treatments did not have the same effect on the two cultivars. For example, all Zn levels influenced the total phenolic concentration of ‘Eleonora’ and ‘Aroma 2’, except the lowest Zn treatment which had no effect on ‘Aroma 2’. Furthermore, the addition of Zn to the nutrient solution overall, had a much greater effect on ‘Eleonora’ plant total phenolic concentration as the latter increased by 47.8% more than that of the ‘Aroma 2’ plants, compared to their respective controls. The CV × Zn interaction for chicoric acid concentration was also significant because while Zn levels up to 37.5 MM appeared to affect the two cultivars similarly, the highest Zn level (50 MM) increased the chicoric acid of ‘Aroma 2’ by 47.4% more than ‘Eleonora’, compared to the control. Regarding the rest of the phenolic acids and depending on the cultivar, some Zn levels had a positive or negative effect on one or the other or both cultivars while others had no effect at all. Specifically, certain levels of Zn  increased the concentration compared to the control of salvianic acid A , caftaric acid , caffeic acid (‘Eleonora’= 42.3%), chlorogenic acid (‘Eleonora’= 13.2%), feruloyl tartaric acid , salvianolic acid K , salvianolic acid A , salvianolic acid L (‘Aroma 2’= 26.2%) and rosmarinic acid . Several Zn treatments had also a negative effect on ‘Aroma 2’ salvianic A (7.8%), caffeic (19%), chlorogenic (21.2%) and rosmarinic (13.1%) acids concentration.Ten phenolic acids were identified in both basil cultivars Table 5.Lactuca sativa L., Brassica oleracea L. var. capitata, Brassica oleracea L. var. italica, and Beta vulgaris L.) when were exposed to Zn biofortified solutions (Oryza sativa and Beta vulgaris L. (–1 of ZnSO4 (–1 dw)  .2, gs, E, SPAD index, and Fv/Fm) and antioxidant activity . The previous response of basil plants after Zn application denotes a typical reaction of plants under heavy metal stress  could have a decisive effect on the Zn concentration and accumulation capacity of the two basil cultivars . The latl stress . Typicall stress . As it tl stress .2). However, adding Zn to the nutrient solution stressed ‘Eleonora’ plants physiology to a greater extent than ‘Aroma 2’ plants when compared to the control. Total chlorophyll concentration, stomatal conductance, and SPAD index of ‘Eleonora’ were reduced to a greater extent than ‘Aroma 2’, by 44.2%, 23.8% and 63.9%, respectively overall Zn treatments. In addition, transpiration rate of ‘Eleonora’ was reduced on average by 19.9% (above 12.5 MM Zn) and Fv/Fm appeared to decrease with increasing Zn concentration while transpiration rate of ‘Aroma 2’ was not affected and Fv/Fm decreased only up to the 25MM Zn level and thereafter remained the same. It must be noted though, that Fv/Fm of both cultivars fell within the optimal range (~0.80) for basil  under Zn surplus. Excessive ROS production under heavy metal stress conditions, such as Zn stress, has been associated with photosynthetic rate limitation and thus plant biomass reduction . On the var used . It was var used . Howeverβ-carotene, are effective chloroplast antioxidants located in close proximity to chlorophylls. At the same time, however, their bioactive oxidized products can induce changes in gene expression that lead to acclimatization to abiotic stress conditions (β-carotene content of ‘Aroma 2’ increased while only β-carotene (at 25 and 50 MM) concentration of ‘Eleonora’ improved when compared to the control. The latter could relate to the increased SPAD index and therefore the photosynthetic efficiency of ‘Aroma 2’ compared to ‘Eleonora’, especially at the higher Zn concentrations. Polyphenolic compounds may have also played an important role in controlling ROS over-production under Zn stress in our study. Chlorophyll concentration reduction under Zn excess and especially the increased production of ROS could have signaled the production of phenolic compounds  and phenolic concentration of ‘Eleonora’ were enhanced and reached higher levels to those of ‘Aroma 2’ control. The latter is very crucial because it could enable us to bio-enhance commercial cultivars whose low concentration of certain compounds beneficial to human health is determined genetically. The health benefits of carotenoids for humans include their general role as antioxidants and the prevention of degenerative macular diseases  both carotenoid (lutein and diseases . Polyphediseases . Kwee anlication .–1 dw) resulted in a yield reduction of less than 10%, but a 15% increase in total phenolics. However, the strong cultivar-dependent response observed in the present study suggests that in biofortification programs for non-hyperaccumulative crops, the varietal choice is crucial in order to maximize the accumulation of this essential micronutrient and minimize yield loss. However, the consumption of biofortified Genovese basil with Zn can increase the intake of Zn by consumers while providing a product enriched concomitantly in valuable phytochemicals such as carotenoids and polyphenols.In this study, we evaluated the potential of producing Genovese basil biofortified with Zn by adopting an agronomic strategy of increasing Zn integration in the nutrient solution . The management of Zn in the floating raft system increased the concentration of Zn in the ‘Aroma 2’ and ‘Eleonora’ basil cultivars while negatively impacting their yield and physiology. However, increasing Zn in the nutrient solution significantly increased antioxidant activity, carotenoid, and polyphenol concentrations. Specifically, the best compromise between yield, phytochemical quality and zinc accumulation in leaves was observed in Aroma 2 biofortified with the highest zinc doze (50 µM). The highest tissue zinc levels in Aroma 2 × 50 .The authors are grateful to Dr. Christophe El-Nakhel for his technical assistance during the experiment.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.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


Both English and Chinese articles about original investigations in humans were searched. Two independent authors extracted the data, and disagreements were resolved by discussion. ReviewManager 5.3 software provided by Cochrane did a meta-analysis of selected randomized controlled trials (RCTs). Quality of experimentation and risk bias were analyzed by the Cochrane Handbook tool. A total of thirteen randomized controlled clinical articles along with 1237 patients were included in the study. Findings of meta-analysis showed that compared with nonacupoint application treatment, the total effective rate of acupoint application treatment is more effective , Z = 6.70, P < 0.00001). Besides, acupoint application can improve patients' lung function, the lung function index FVC , Z = 8.40, P < 0.00001), FEV1 , Z = 5.86, P < 0.00001), FEV1/FVC (%) , Z = 2.97, P = 0.003), FEV1 (%) , Z = 27.44, P < 0.00001), and PEF (day) , Z = 12.40, P < 0.00001) of patients treated by acupoint application therapy were increased. Moreover, acupoint application might lower the level of immunoglobulin E , Z = 11.93, P < 0.00001) and EOS , Z = 2.77, P = 0.006). The LCQ (Leicester cough questionnaire) total score of CVA patients was also increased , Z = 5.98, P < 0.00001). Acupoint application therapy is effective in controlling symptoms of CVA. It also has a positive effect in improving lung function and life quality of patients. It can reduce the eosinophil levels and peripheral blood IgE levels of patients as well.Acupoint application has been used in China to treat various illnesses for ages. In cough variant asthma (CVA), the main clinical sign is episodic night cough. Acupoint application therapy of traditional Chinese medicine is an effective procedure to treat cough variant asthma. The current study is designed to systematically assess the effectiveness of acupoint application therapy in traditional medicine for patients with cough variant asthma. The comprehensive computer retrieval related to comparison between acupoint application and nonacupoint application therapy for cough variant asthma was carried out in various databases ( Cough variant asthma (CVA) is a particular form with typical common cold symptoms and dry, nonproductive cough. Other manifestations, like dyspnea or gasping, are not generally observed; however, there could be episodic night cough which can be alleviated by a bronchodilator . The symIn addition, the prevalence of cough variant asthma is still prominent due to air pollution , smokingCurrent treatments for CVA are generally the same as ordinary respiratory illness, along with bronchodilators, glucocorticoid drugs, antihistamines, and leukotriene receptor antagonists . AlthougAcupoint application (AP) is a traditional Chinese medicine (TCM) method with a long history. The main steps of this treatment are grinding the herbs into powder and turning them into herbal patches, directly sticking to acupoints or affected areas to treat chronic cough. Studies have shown that acupoint application can affect the level of immunoglobulin and eosinophils in patients with CVA, regulate the proportion of lymphocytes, and influence the proportion of some cytokines, such as TGF, TNF, and IF, to control the symptoms of CVA and achieve long-term relief –8.Recently, there have been many studies demonstrating AP's positive results for curing CVA . Howeverhttps://pubmed.ncbi.nlm.nih.gov/), EMBASE (https://www.embase.com/landing?status=grey), Web of Science (https://mjl.clarivate.com/search-results), the Cochrane Library (https://www.cochranelibrary.com/), Chinese Journal Full-Text Database (CNKI) (http://kns55.en.eastview.com/kns55/brief/result.aspx?dbPrefix=CJFD), Database of Chinese Sci-Tech Periodicals (VIP) , “Wanfang” Database (http://www.wanfangdata.com/), and China Biology Medicine Disc (CBM). The following keywords were used: “stimulating acupoints,” “acupoint sticking,” “traditional Chinese medicine,” “TCM,” “acupoint,” “CVA,” “cough variant asthma,” “cough type asthma,” “cough-variant asthma,” “randomized controlled trial,” “random,” “control and trial,” and “RCT.”We systematically searched the literature for the formation of every database to July 4, 2021. Databases include PubMed  OR Traditional Chinese Medicine[MeSH Terms]) OR Chinese Traditional Medicine[MeSH Terms]) OR acupuncture[Abstract]) OR moxibustion[Abstract]) OR “auricular points plaster therapy”[Abstract]) OR “acupoint sticking”[Abstract])) AND “Cough Type Asthma”[Abstract]) OR “Cough variant asthma”[Abstract]) OR “Cough-Type Asthma”[Abstract]) OR “Cough-variant asthma”[Abstract]).#1 “auricular points plaster therapy”[Abstract/Title].#2 “acupoint sticking”[Abstract/Title]).#3 Traditional Chinese Medicine[MeSH Terms].#4 “ #1 OR #2 OR #3 ”.#5 “Cough Type Asthma”[Abstract/Title].#6 “Cough variant asthma”[Abstract/Title].#7 “Cough-variant asthma”[Abstract/Title].#8 “CVA” [Abstract/Title].#9 “ #5 OR #6 OR #7 OR #8”.#10 “Randomized controlled trial”[Abstract/Title].#11 “Random”[Abstract/Title].#12 “Control”[Abstract/Title].#13 “Trial”[Abstract/Title].#14 “ #10 OR #11 OR #12 OR #13”.#15 “ #4 AND #9 AND 14”.http://www.who.int/ictrp/en/) were also explored.Moreover, to include all the possible information, incomplete and finished experiments on the Chinese Scientific Experiments Register (update to July 2021) and World Health Organization ICTRP . These parameters were studied and included in the analysis.From the studied sections, participants of those studied articles were also screened. The following features of the study participants, age limit, gender, disease identification, other signs, and treatment count, as well as control samples, key features of treatment and control groups, and total completed experiments as well as incomplete or withdrawn, were taken for further analysis.In interventions, site for acupoint application, time of interference, and noninterference were focused.From the results section of the literature, total effective rate, forced expiratory volume (FVC), forced expiratory volume in one second, FEV1/FVC (%), FEV1/predication (FEV1/pre), peak expiratory flow (PEF), EOS count per milliliter in peripheral blood, IgE level per milliliter in the peripheral blood, and LCQ total score were taken for further study and analysis.Evaluation of standard of research was done by ReviewManager 5.3 software risk bias assessment tool equipped from Cochrane Collaboration: (a) to generate the randomized data, (b) concealing the allocation, (c) blinding of research individuals, (d) blinding of the evaluation of results, (e) no proper information retrieved, (f) prediction of specific results, and (g) different partialities. Each group was regarded as “high risk of bias” and “low risk of bias”/“unclear risk of bias.”We used the Cochrane ReviewManager 5.3 software for meta-analysis and assessment of reviewed data. Dichotomous data were displayed as odds ratio/risk ratio having 95% confidence intervals that predict the chance of risk or relative risk. Continuous variable dataset assessment was done by MD odds ratio and 95% confidence intervals (CIs). Key point of evaluation is the research volunteers.P value: if P value was less than 0.10, this showed statistically significant results. I2 statistic was applied as guided by the Cochrane Handbook for Systematic Reviews of Interventions. IfI-square (I2) was used as a statistical method to assess data heterogeneity, the value of I2 < 40% was indicative of less heterogeneity, whereas more than 75% indicated significant heterogeneity in experimentation. The funnel plot visually analyzed the “risk of reporting” bias.Assessment of the experiments was done for clinical heterogeneity , diversity in methodology , and statistical diversity. The chi-square test was applied with a I2 > 50%, the random-effect model (REM) was applied for assessment.Sensitivity analysis was done as described below: assessment of outcomes of two statistical models, i.e., random-effect model (REM) and fixed model, were applied and compared. If A total of 534 records were analyzed after excluding the duplicated data. This data was further meta-analyzed, and all the information related to the search for knowledge and protocol selection for research is presented in Out of the screened articles, thirteen articles were disclosed in 2014-2020 from nine provinces in China about AP in CVA. A total of 1237 volunteers participated in these researches, aged of 4-65 years. In previously published articles, the test groups consisted of patients who had undergone AP along with different treatments. In the control group, no AP was applied, and only Western or traditional Chinese medicines were used for the treatment of CVA.The acupoint application test and control samples had 621 and 616 cases, respectively. Key features of all elected research are mentioned in Assessment of “risk of bias” is presented in 2 = 11.37, P = 0.33, and I2 = 12%, and the fixed-effect model was applied for evaluation . The funnel plot revealed a uniform plot on both sides (left and right) and stacked on the upper side, showing a certain risk of bias, as shown in The sum of eleven types of research , 12–21 ialuation . Total eZ = 8.40, P < 0.00001)  Z = 5.86, P < 0.00001). These findings suggested that AP has more ability to improve the pulmonary function index FEV1 compared to the control group , and a subgroup analysis was performed. In the subgroup of patients younger than 7, the subgroup evaluation of four researchers reported a statistically small difference , Z = 8.00, P < 0.00001). In the subgroup of patients older than 7, the subgroup analysis showed a statistically significant difference , Z = 6.93, P < 0.00001).Five articles , 18, 22 ol group . A test Z = 2.97, P = 0.003). This suggests that acupoint application improves pulmonary function index FEV1/FVC (%) better than the control group. Heterogeneity among researchers  and subgroup analysis were performed. For the subgroup of patients whose intervention period is less than 30 days, subgroup evaluation in the 180 patients resulted in a statistically significant difference , Z = 5.65, P < 0.00001). Statistically significant results were found in those patients (n = 120) whose intervention period was more than 30 days and less than 6 months , Z = 12.00, P < 0.00001), whereas 120 patients were found with statistically significant results whose intervention period was more than 6 months , Z = 13.72, P < 0.00001)  .Z = 27.44, P < 0.00001). The difference between the three subgroups was significantly small (I2 = 0%)  , I2 = 0%) .Z = 12.40, P < 0.00001) which demonstrated that AP treatment is more effective in improving the pulmonary function index PEF compared to the control group , which reported that acupoint application treatment could better decrease the peripheral blood IgE level of CVA patients as compared to control samples , the cough variant asthma measure for acupoint application therapy (I2 = 66%). The random-effect analysis algorithm was applied to analyze two groups of samples , Z = 2.77, P = 0.006), indicating that acupoint application treatment could better decrease the peripheral blood EOS count of CVA patients as compared to control samples  , evaluating acupoint application treatment which might lead to better increase of the LCQ score of CVA patients compared with the control group  . The fixol group .It is widely believed that CVA is regarded as a particular form of respiratory illness with a histopathological process, just like common asthma. These include hyperresponsiveness (BHR), eosinophilic airway inflammation, and airway remodeling . Variousβ2 receptor agonists, inhaled anticholinergic drugs, and short-acting theophylline. These drugs are very effective in alleviating symptoms, reducing airway inflammation, and improving quality of life. However, adverse reactions of these drugs have also been observed. Glucocorticoids may cause hoarseness of voice and oral candida infection. The use of β receptor agonists may lead to sympathetic nerve excitation and accelerated heart rate, resulting in palpitations, chest pain, and other symptoms. So, it is advised to use these drugs only in emergencies and for the shorter period of time. Acupoint application is a nice alternative to these drugs [Asthma can be treated by inhaled glucocorticoids and leukotriene modulator drugs, etc. These drugs are categorized into a control group and need to be taken for a long period for the therapy to be effective, whereas other drugs include palliative drugs which include short-acting 13se drugs .Acupoint sticking therapy has a long history in traditional Chinese medicine. The main steps of this treatment are as follows. First, a variety of herbs were ground into a powder. Second, adhesive materials such as ginger water were prepared. Next, mix the powder with the adhesive to make a pulp salve that may look like “caking agent” and put it on certain acupuncture points of the body. For treating cough variant asthma, we often choose the “TianTu” (RN22), “DaZhui”(DU14), “FeiShu”(BL13), and “ShanZhong” RN17) acupoints. Acupoint application allows drugs to be absorbed directly through the skin into capillaries without the need for liver metabolism, which preserves the biological activity of some drugs  acupoint.Acupoint application has an advantage over other treatment therapies in treating asthma as acupoint stimulation promotes flow of blood to dispel pathogenic factors. This can stimulate the body's immunity and reduce allergic states . But stiFindings of this meta-analysis revealed that the rate of effectiveness, lung function index , FEV1 (%), PEF (day)) of the CVA sample showed more significant values as compared to the control group, whereas IgE and peripheral blood EOS count showed lower values when compared with control samples. This suggested that AP for cure of CVA has better efficacy than the other drug treatments.The main advantage of this study is that we conducted a meta-analysis of 13 RCTs involving 1237 participants. Compared to previous systematic reviews of acupoint application for CVA , this stHowever, this study still has some limitations. The foremost limitation is that, although the treatment of CVA by acupoint application is frequently used, random clinical controlled studies are usually single-center studies with a small sample size. There are problems such as having no recognized standard for efficacy evaluation and clinical heterogeneity. These problems suggest the need for high-quality clinical research methods in treating CVA by acupoint application, including correct randomization, double-blind, and allocation concealment methods, as well as large-scale multicenter studies. Second, since the acupoint application requires the application operating on the patient, and the herbs have a special smell, it is impossible to blind the patients during the operation in all employed research. Therefore, based on risk-of-bias assessment software provided by the Cochrane Organization, the “blinding of participants and personnel” in whole reports was evaluated as “high risk.” Third, the language of retrieval in the present research was in Chinese and English, and the literature was only from 8 databases. Besides, all the reports used in the meta-analysis were in Chinese, and all the experiments were conducted in China, limiting the present results' specifications because of sample features. Fourth, due to the complexity of acupoint application, this study mainly focused on the treating method of acupoint application but did not explore the influence of different acupoint selections and the type of herbal medicine on treating effectiveness.The current study concluded that acupoint application is better for the CVA treatment than the control group, which was treated with other traditional medicines. Moreover, it was observed that AP improved respiration and chronic airway inflammation by reducing eosinophil levels and peripheral blood IgE levels.


FTO (rs9939609), FABP2 (rs1799883), and LEP (rs2167270), LEPR (rs1137101), and MC4R (rs17782313) polymorphisms and obesity-related parameters. Unrelated Caucasian males (n = 165) were recruited. All participants had similar physical activity levels. The participants were divided into two groups depending on their body mass index (BMI) and fat mass index (FMI). All samples were genotyped using real-time polymerase chain reaction . When tested individually, only one statistically significant result was found. The FTO A/T polymorphism was significantly associated with FMI (p = 0.01). The chance of having increased FMI was >2-fold higher for the FTO A allele carriers (p < 0.01). Gene–gene interaction analyses showed the additional influence of all investigated genes on BMI and FMI. In summary, it was demonstrated that harboring the FTO A allele might be a risk factor for elevated fat mass. Additionally, this study confirmed that all five polymorphisms are involved in the development of common obesity in the studied population and the genetic risk of obesity is linked to the accumulation of numerous variants.Obesity is a complex multifactorial abnormality that has a well-confirmed genetic basis. However, the problem still lies in identifying the polymorphisms linked to body mass and composition. Therefore, this study aimed to analyze associations between  Obesity is defined as excessive body weight gain due to an increased accumulation of body fat that presents a leading cause of the largest public health problems , since nObesity is a complex multifactorial disease that has a well-confirmed strong genetic basis but needs behavioral, developmental, and/or environmental influences to develop ,9,10. SeFTO), fatty acid-binding protein 2 (FABP2), leptin (LEP), leptin receptor (LEPR), and melanocortin-4 receptor (MC4R), which are characterized in Although studies of the common obesity genetics were determined by genome-wide association studies (GWASs), this stage was set by research on monogenic obesity, which emphasized that the leptin–melanocortin signaling pathway is the major regulator of food intake. Several genes involved in the development of monogenic obesity are in or near loci subsequently linked by GWASs with obesity-related traits. To date, over 600 genes and chromosomal regions have been associated with the regulation of body weight and composition ,13. The FTO (rs9939609), FABP2 (rs1799883), LEP (rs2167270), LEPR (rs1137101), and MC4R (rs17782313) polymorphisms and obesity-related traits. Therefore, we studied individually and in gene–gene interaction models the alleles and genotypes distribution in a group of physically active Caucasian men measured for selected body mass and body composition traits.The above-mentioned findings highlight the selected genes’ association with body mass and body composition parameters. However, the results are inconsistent and the interactions between these candidate genes are still unknown. Thus, this study aimed to examine the associations between One hundred and sixty-five unrelated military professionals were selected for this study. All participants were aged 19–54 years, ancestrally fitted , and they represented similar physical activity levels. The experimental protocols used in this study were conducted in accordance with the World Medical Association’s Declaration of Helsinki and were positively verified by the Ethics Committee of the Military Institute of Hygiene and Epidemiology (no. 1/XXI /2016). The participants received an information sheet regarding the research details, aim of the study and procedures applied, as well as potential risks and benefits associated with their participation. All volunteers gave written, informed consent for the genotyping, and they were informed that the study will be anonymous and the results would be private.2), height (cm), weight (kg), fat mass index  [Height was measured using a portable stadiometer (without shoes) . Body composition (including fat %) and body weight were measured using bioelectrical impedance analysis (BIA) using the TANITA MC-780 machine  with an accuracy of 0.1 kg according to the procedure specified in the instruction manual . All measurements were performed according to the procedure specified in the instruction manual and without any metal objects. The following parameters were noted: BMI  , visceran = 165) were divided into two groups depending on their BMI . The control group  comprised people with BMI between 20.0 and 25.0, while the overweight group  had a BMI of ≥25.0. They were also divided into two groups depending on their FMI . FMI values between 3 and 6 were classified as normal fat mass; FMI lower than 3 —fat deficit; FMI higher than 6—excess fat. Participants whose FMI values were 6 and lower were classified into the CONFMI group (n = 124), while those whose FMI values were higher than 6 were grouped into the OVERFMI group (n = 41). Statistically significant differences between participants in both groups were observed for parameters BMI (kg/m2), age (years), weight (kg), FMI (kg/m2), VTI (level) and Fat(%) . No statistically significant difference was shown in the parameter height (cm)   . DetaileThe buccal cells of the participants were collected using swabs . Genomic DNA was extracted from the donated buccal cells using a High Pure PCR Template Preparation Kit  according to the manufacturer’s protocols. DNA samples of good quality and quantity were stored at -20 °C. The exclusion criteria were: failure in DNA extraction; DNA degradation; abnormal gene detection results; incomplete basic information. All samples were genotyped in duplicate, using TaqMan Pre-De-signed SNP Genotyping Assays, which are given in https://cran.r-project.org (accessed on 20 September 2021)). Anthropometric data are shown as mean values ± standard deviation and differences among experimental groups were analyzed with Student’s t-test which was statistically significant when p < 0.05. To check the compliance of the variables with the normal distribution, the Shapiro–Wilk test was used and Levene’s test was used for verification of the homogeneity of variance. Single-locus analysis was performed considering four genetic models  and was calculated with the SNPassoc package for R. The models were constructed concerning the minor allele and were checked adjusted by age as a potential factor influencing the result. FDR-adjusted p-values were calculated with the fdrtool package for R. An odds ratio (OR) was used as a measure of association between an exposure and an outcome and to determine whether a particular genotype is a risk factor for being overweight. The Akaike information criterion (AIC) was used to evaluate how well a model fits the data. A genetic model-free Multifactor dimensionality reduction (MDR) was used to detect the influence of the common effect of gene × gene interactions on BMI and FMI, and it was calculated with MDR3.0.2 (http://sourceforge.net/projects/mdr/ (accessed on 20 September 2021)); chi-square test was used for checking the statistical significance of the model. Balance accuracy was used as the evaluation measure to rank potential models and cross-validation consistency was used to choose the best models. The association of single alleles with BMI was calculated with Pearson’s chi-squared test with the STAT package for R. The level of statistical significance was set at the level of p < 0.05. Genotype frequencies were analyzed using Fisher’s exact test with the STAT package for R.All statistical analyses were performed using the program R , CONBMI group , OVERFMI , CONFMI , and the case–control group , FABP2 (rs1799883), LEP (rs2167270), LEPR (rs1137101), MC4R (rs17782313) and the BMI value exceeding 25. All divisions were checked under four genetic models: codominant, dominant, recessive, and overdominant, p-values were between 0.11 and 0.98. The influence of single alleles on BMI division was also checked and no significant association was found (No significant association was found between as found .FTO gene polymorphism (rs9939609) was significantly associated with FMI exceeding 6 (FMI for the combination AA was over 4.7 times greater than for the combination TT in the codominant model (Fisher’s exact test p = 0.01). The chance of being OVERFMI for the combination AT-AA was >2.7 times higher than for the combination TT in the dominant model (Fisher’s exact test p = 0.02). The chance of being OVERFMI for the combination AA was >2.8 times higher than for the combination TT-AT in the codominant model (Fisher’s exact test p = 0.02). Moreover, the chance of being OVERFMI was >2.0-fold higher for A allele with Pearson’s chi-squared test p-value < 0.01. The model was supplemented with age as a potential factor influencing the result, because of statistical differences shown between groups for this variable (The eeding 6 . An assovariable .FTO (rs9939609) and LEPR (rs1137101), indicating a potential gene–gene interaction between these two genes. For BMI division, when genotypes AT × AA, TT × AA, TT × AG  appear the model sorts the observations to join the CONBMI group with a higher probability than joining the OVERBMI group (p = 0.02). For FMI division when the genotypes AT × GG, TT × AA, and TT × AG  appear, the model sorts the observations to join the CONFMI group with a higher probability than joining the OVERFMI group , LEPR (rs1137101), and MC4R (rs17782313). The genotypes GG × AA × TT, GG × AG × TC, GG × AG × CC, GG × GG × TT, AA × AA × TT, AA × AA × TC, AA × AG × TT, AA × AG × CC, AG × AA × TT, AG × AG × TT, and AG × GG × CC  were selected by an algorithm to join the CONBMI group (p < 0.01). For FMI division, the best three-locus model included the genes FTO (rs9939609), LEPR (rs1137101), and MC4R (rs17782313). The genotypes AT × GG × AA, AT × AG × GG, TT × GG × AG, TT × AG × AA, TT × AG × AG, and TT × AA × AG  were selected by an algorithm to join the control group with a higher probability (p < 0.01). When the genotypes TT × GG × GG, and TT × AA × AA  appeared, the model sorted the observations to join the CONFMI group (p < 0.01). In all divisions in the four-locus and five-locus models when genotype was chosen to join the control group in FTO (rs9939609), only TT and AT genotypes appeared (The best chosen three-locus model for BMI division involved the genes appeared .FTO (rs9939609), FABP2 (rs1799883), LEP (rs2167270), LEPR (rs1137101), and MC4R (rs17782313) polymorphisms in a group of Caucasian men, who were divided into groups depending on their BMI and FMI. When tested individually, our statistical analyses showed that harboring the specific FTO genotype might be associated with FMI, which measures relative fat content. This association was found for the codominant (A/A vs. T/T), dominant (A/T-A/A vs. T/T), and also for the recessive genetic model (A/A vs. T/T-A/T). The chance of having an increased FMI was over two times higher for the FTO A allele carriers. This observation constitutes the first important finding of the present study, implying that harboring this specific allele is unfavorable for some individuals. The carriers of the A allele might have an increased accumulation of body fat and a higher risk of many obesity-related disorders. In the present study, we examined the allele and genotype distributions of the FTO first intron was strongly related to body mass and composition parameters and predisposes to overweight and obesity in children, teenagers, and adults [FTO variants, especially the common A/T polymorphism (rs9939609), are significantly linked to obesity-related traits, e.g., BMI, FMI, body fat percentage, hip circumference, cardiometabolic traits, and many obesity-related medical problems. These associations are found across various ethnic populations and different age groups [FTO gene can influence athlete status in a study involving 196 elite swimmers and 379 control participants, who were all Caucasians. They found that harboring the T allele might be favorable for achieving success in sports such as swimming [FTO (rs9939609) polymorphism is a candidate marker for affecting body mass and body composition parameters in the Caucasian population.In 2007, three independent groups demonstrated that a cluster of polymorphisms in the d adults ,26,27. Se groups ,29. The e groups . Conseque groups . In a prswimming . These rFTO polymorphism and body mass and composition parameters have been revealed. The research has indicated that these associations are mediated through their functional interactions with distal surrounding genes. The first intron of the FTO gene contains a binding site for the transcriptional factor—cut-like homeobox 1 (CUX1). Through controlling retinitis pigmentosa GTPase regulator interacting protein 1 (RPGRIP1L) expression, it interacts with the leptin receptor [FTO intron also includes an enhancer sequence, which interacts with the iroquois homeobox 3 (IRX3) promoter region, and thus the FTO SNPs regulate IRX3 expression in the human brain. The IRX3 relationship with obesity and the process of browning in adipose cells has been described [FTO gene is closely related to the regulation of levels of growth hormone and insulin-like growth factor I (IGF-1). IGF-1 is a crucial hormone in the development of metabolic syndrome, due to its influence on lipid and carbohydrate metabolism [A few potential biological mechanisms underlying the relationship between the receptor ,31. The receptor . Additioescribed ,31,32. Ctabolism .FTO × LEPR interaction—both the BMI and FMI division genotypes, TT × AA and TT × AG, were associated with the absence of overweight. The same result showed the genotypes AT × AA with BMI division and AT × GG for FMI division. For three-locus model genotypes, GG × AA × TT, GG × AG × TC, GG × AG × CC, GG × GG × TT, AA × AA × TT, AA × AA × TC, AA × AG × TT, AA × AG × CC, AG × AA × TT, AG × AG × TT, and AG × GG × CC (LEP × LEPR × MC4R) showed a link to the lack of overweight in BMI division. The same association was shown for genotypes AT × GG × AA, AT × AG × GG, TT × GG × AG, TT × AG × AA, TT × AG × AG, and TT × AA × AG (FTO × LEPR × MC4R) in FMI division. For four-locus and five-locus models, only the genotypes TT and AT in FTO present when there was an association with the absence of overweight shown (both for BMI and FMI). In all models, when FTO was included, the genotypes TT and AT were linked with a lack of overweight, confirming that harboring the FTO T allele might be favorable for some individuals.When the results obtained in our study were incorporated into the complex gene–gene interaction analysis, the novel finding was that all five studied polymorphisms are involved in the formation of obesity-related traits in the Caucasian population. These results imply that some individuals might benefit from carrying some combinations of genotypes. It was shown that for the two-locus model—FTO genotype and FMI, the gene–gene interaction analysis revealed numerous links between the genotypes of studied genes and obesity-related traits. This observation confirmed that the genetic risk of obesity is connected with the accumulation of numerous variants; thus, methods based on numerous SNPs are more informative than methods based on a single polymorphism. Cole et al. showed that the analysis of gene–gene interactions is a potential source of unexplained heritability, a significant focus of research into complex traits, including obesity, which involves a complex interaction between several genetic variants. Such polygenic traits frequently require etiologies in which complicated biological relations within different tissues, pathways, and networks underlie the trait development [Although the analysis of individual SNPs showed only one association between harboring the specific elopment . Studyinelopment , which iBDNF, ETV5, FAIM2, FTO, GNPDA2, KCTD15, MC4R, MTCH2, NEGR1, SEC16B, SH2B1, and TMEM18). The authors underlined that the used methodology made it possible to reveal the background of interactions between genes known to influence BMI. They characterized the complicated interactions, emphasized new roles of the genes and highlight the involvement of regulatory frameworks in the development of obesity; e.g., rs17066891 in MC4R was identified as having the strongest main effect within this network, rs9940128 in FTO was identified as having the second strongest main effect in the network, and rs4358154 in TMEM18 had the highest score for all measures which highlights the potentially significant role of this variant in the context of obesity [MC4R rs17782313, BDNF rs6265, FTO rs1421085, TMEM18 rs7561317, and NEGR1 rs2815752). Surprisingly, the gene–gene interaction was not found to significantly influence any obesity-related anthropometric phenotype, such as BMI or body fat percentage [Although numerous studies refer to the analysis of individual SNPs, which were selected for the present study, the literature on gene–gene interaction analyses is scarce. Therefore, the obtained results cannot be discussed with direct comparisons to other studies. In the study including 2386 individuals, De et al. analyzed interactions between twelve genetic variants robustly associated with obesity  indicated that fasting insulin in women shows stronger genetic correlations than in men with waist-to-hip ratio and anorexia nervosa . UnfortuFTO gene, when tested individually, and all selected genes , when tested in gene–gene interactions, can modify body weight and composition parameters (BMI and FMI). We showed that harboring the FTO A allele might be associated with an over two times increased FMI. This observation constitutes the first important finding of the study, implying that harboring this specific allele is unfavorable for some individuals. The carriers of the A allele might have an increased accumulation of body fat and a higher risk of many obesity-related disorders. The results suggested that the FTO (rs9939609) polymorphism is a candidate marker for affecting body mass and body composition parameters in the Caucasian population. From the gene–gene interaction analysis, we established the second novel finding. Namely, all five studied polymorphisms are involved in the formation of obesity-related traits in the Caucasian population. The results imply that some individuals might benefit from carrying some combinations of genotypes , as regards the BMI and FMI. Additionally, the gene–gene interaction analysis confirmed that harboring the FTO T allele might be favorable for some individuals. This observation confirmed that the genetic risk of obesity is connected with the accumulation of numerous variants; thus, methods based on numerous SNPs are more informative than methods based on a single polymorphism. Understanding the genetics of obesity can extend our knowledge of diet individualization and exercise programs. It is crucial for the prevention of obesity-related diseases, supplementation and medical care.The results of the present study suggest that the