Objective: To explore the detective value of magnetically controlled robotic capsule endoscopy (MCRCE) in patients with suspected small intestinal disease.Patients and Methods: In total, 1,802 patients with suspected small intestinal disease and negative gastroenteroscopy from Shanghai Jiao Tong University Affiliated Sixth People's Hospital were examined with MCRCE, and the data were collected for further analysis.Results: Among the 1,802 patients who were examined with MCRCE, 974 were diagnosed with small intestinal disease, reaching a positive detection rate of 54.1%. The five most common conditions that were detected include non-specific enteritis in 722 cases (40.1%), small intestinal ulcers in 87 cases (4.8%), abnormal small bowel evacuation in 45 cases (2.5%), small intestinal bleeding in 33 cases (1.8%), and small intestinal yellow spots in 31 cases (1.7%). The running time of the capsules in the small intestine ranged from 85–437 min, with an average of 210.24 ± 89.08 min. No complications, such as intestinal obstruction or capsule retention, were observed in all patients.Conclusion: MCRCE is a safe and non-invasive endoscopic examination with a highly accurate detection rate for small intestinal diseases. With a total length of 5–7 m in adults, the small intestine is the longest part of the alimentary canal. Diseases of the small intestine are important components of digestive tract diseases. These diseases are clinically present as gastrointestinal bleeding, abdominal pain, diarrhea, and constipation. Lesions of the small intestine mainly consist of benign ulcers, tumors , chronic inflammation, vascular malformations, polyp, and diverticula –3. The iPrevious studies showed that the detection methods for gastroenterologic diseases include computerized tomography (CT), enterography, digital subtraction angiography, and balloon-assisted enteroscopy. However, these diagnostic methods cannot be widely used for the small intestine because of their low sensitivity, complexity of operation, and invasiveness , 5. MoreThe second generation of magnetically controlled robotic capsule endoscopy (MCRCE) is a new and advanced clinical detection technology in China with a high detection rate for small lesions in the small intestinal tract using a non-invasive and painless procedure . In thisPatients from the gastroenterology outpatient and inpatient department of Shanghai Jiao Tong University Affiliated Sixth People's Hospital, who were suspected of having small bowel disease, between March 2016 and May 2019 were included in this study . All patMCRCE was developed by Shanghai Ankon Medical Technology Co., Ltd. and Wuhan Ankon Photoelectric Technology Co., Ltd. . The NaviCam MCRCE  comprises a capsule control system, a capsule endoscope, a portable recorder, a capsule positioner, and a display software. The control system includes the testbed, magnetic head, rotary table, and control table. The main specifications of the NaviCam magnetic capsule endoscopy are as follows: size of 27.0 × 11.8 mm, visual angle of 140 ± 14 degrees, shooting frame rate in the stomach or small bowel mode of two pictures per second, working time of 10 14 h, and net weight of 2 g . The batAs shown in Images were reviewed independently by two experienced senior physicians who agreed upon the final diagnosis. Non-specific enteritis was defined as an inflammation located in any one of the duodenal, jejunal, and/or ileal parts of the small bowel. Inflammation was defined as having the following manifestations: (i) mucosal lesions (including erosions); (ii) mucosal changes, such as edema and prominent mucosal folds; (iii) changes in the villi, such as flat mucosa and coarsened villi; and (iv) lymphangiectasias/lymphocellular infiltrates , 7. The Statistical analysis was performed using SPSS 22.0 . Data that were normally distributed are expressed as mean ± standard deviation, and other recorded data were expressed in terms of the number of cases and percentages.In total, 1,802 patients suspected of having intestinal disease were enrolled in the study. These included 811 men and 991 women, aged 14–95 years, with a mean age of 50.10 ± 16.26 years. Abdominal pain and discomfort were observed in 630 (35%) and 290 cases (16.1%), respectively. Other clinical symptoms of the patients included in the study are presented in During the study's timeline, 1,960 of the 1,966 patients (99.69%) successfully completed the examination. Six of the subjects were lost to follow up. Among the 1,960 participants, 158 were excluded due to the lack of etiological data, while 1,802 were suspected of having small bowel disease. Among these 1,802 participants, 828 had normal findings in their small intestines, while the remaining 974 were identified to have small bowel disease . All patThe operator controlled the capsule into the duodenum in 255 cases (14.2%), while it was naturally transported into the duodenum in the remaining 1,547 cases (85.9%). The image quality and clarity were ideal in most subjects; however, observation of some areas of the small intestinal mucosa is limited by bile and other small bowel contents and secretions. The running time of the capsules in the small intestine was 85–437 min, with an average of 210.24 ± 89.08 min.The MCRCE procedure could detect various small bowel conditions in a total of 974 participants, the summary of which is presented in The anatomical structure of the small intestine is so complex that conventional endoscopy cannot fully examine the entire small intestine. Enteroscopy is a commonly used procedure for diagnosing small bowel diseases. As an invasive examination, it requires a high degree of technical operating skills and a long operating time for physicians. Moreover, enteroscopy is associated with perforation, pancreatitis, and bleeding , 9. In cIn 2014, we began to use MCRCE to detect small intestinal disease, providing a non-invasive examination of the whole small intestine. It overcame the restriction of conducting endoscopy for patients with history of severe cardiopulmonary or spinal diseases, and endoscopy is known to be contraindicated in these cases. MCRCE can provide patients, who have absolute or relative traditional endoscopic contraindications, with timely endoscopy, early diagnosis, and treatment. MCRCE not only provides painless and non-invasive examinations, but also prevents cross-infection, narcotic drug risk, and other adverse effects in patients. All these factors make MCRCE a safer and more acceptable examination technology for small bowels .In this study, the majority of patients successfully completed the examination. Six of the subjects were lost to follow up, and 158 of the subjects were excluded due to inadequate etiological data . RelatedFurther, MCRCE helped detect small intestinal lesions in 974 of the 1,802 patients with gastrointestinal symptoms. Therefore, the positive detection rate of 54.1% suggests a high incidence of small bowel disease in patients with chronic abdominal pain, abdominal distension, diarrhea, and unknown bleeding. MCRCE is highly clinically valuable in the detection of small intestinal diseases, consistent with the findings of previous studies , 14, 15.Small intestinal bleeding emerges from between the suspensory ligament of the duodenum and the Bauhin's valve, accounting for 5% of all gastrointestinal bleeding (GIB) and 80% of obscure GIB cases . Common However, the main limitations of MCRCE include: (i) different observed areas depending on the movement trajectory of the capsule; (ii) the long time of processing and reporting ; (iii) aThis study suggests that MCRCE is a potential primary choice for the examination of small bowel diseases. With the progress of science and technology, capsule endoscopes with more comprehensive functions are being developed, such as biopsy capsules, hemostatic capsules, and drug transport capsules –24. We bMCRCE is a safe and non-invasive technology, with a highly accurate detection rate in diagnosing small intestine diseases, which is significantly valuable for clinical application in patients with suspected small intestine diseases.The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation.The studies involving human participants were reviewed and approved by the clinical research was approved by the medical device ethics committee of Shanghai Jiao Tong University Affiliated Sixth People's Hospital. The patients/participants provided their written informed consent to participate in this study.J-SZ and JZ supervised the study. X-YC and JZ oversaw the study design. X-YC, WD, RL, H-NF, Y-CY, MC, and H-WQ performed the procedures. X-YC, WD, and JZ performed the data analysis. X-YC prepared the manuscript while JZ and J-SZ revised the manuscript. All authors contributed to the article and approved the submitted version.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.


Obesity is a pandemic condition of complex etiology, resulting from the increasing exposition to obesogenic environmental factors combined with genetic susceptibility. In the past two decades, advances in genetic research identified variants of the leptin-melanocortin pathway coding for genes, which are related to the potentiation of satiety and hunger, immune system, and fertility. Here, we review cases of congenital leptin deficiency and the possible beneficial effects of leptin replacement therapy. In summary, the cases presented here show clinical phenotypes of disrupted bodily energy homeostasis, biochemical and hormonal disorders, and abnormal immune response. Some phenotypes can be partially reversed by exogenous administration of leptin. With this review, we aim to contribute to the understanding of leptin gene mutations as targets for obesity diagnostics and treatment strategies. The equilibrium between the amount of energy intake and expenditure is the key to fat tissue homeostasis. Such an equation is not that simple as it seems. Obesity is a complex disease defined as abnormal or excessive adiposity that may cause different levels of health impairment. In the last two decades, obesity has shown an expressive growth rate, assuming global epidemic proportions . To dateThe increase in the global burden of obesity has been attributed to the interplay between distinct factors, such as environmental factors, e.g., urban progress that generated daily facilitating mechanisms, more passive forms of entertainment, and reduced physical activity, leading to a spread of sedentary lifestyle . ConcomiBoth maternal and paternal BMI are correlated with offspring BMI, supporting family history of overweight/obesity as a risk factor for childhood overweight/obesity . Modern Over the last 20 years, a large number of different rare variants were described in genes encoding proteins within the leptin-melanocortin pathway, involved in hypothalamus cell differentiation, appetite modulation, and energetic metabolism regulation. These may cause early-onset obesity, hyperphagia, and endocrine abnormalities. Mendelian nonsyndromic obesity, alias monogenic nonsyndromic obesity is associated with only 5% of obesity cases in the population. Though rare, it is the most severe form of the disease , 14.LEP was the first gene associated with nonsyndromic monogenic obesity in 1997 . LRb is the longest form (1162Aa) from a family of six LR isoforms (LRa-LRf), differing only in the intracellular tail length . LRb is Tyr985 poses a binding site to the src-homology 2 domain protein (SHP2) that modulates extracellular signal-regulated kinase (ERK) 1/2 pathways. JAK2-ERK 1/2 activation was shown to diminish appetite, decrease body weight, and modulate the thermogenic sympathetic outflow . Tyr1077The STAT3 transcription factor also inhibits AgRP and NPY gene expression and upregulates POMC –44. FurtDuring fasting and decreased energy availability, serum leptin levels are diminished, and the ghrelin hormone is secreted by the stomach. It binds to its receptor at orexigenic neurons of the arcuate nucleus, upregulating NPY and AgRP expression. These neuropeptides act as melanocortin receptor antagonists and promote increased food intake and decreased energy expenditure , 56.ob) gene, Lep and its human homolog gene (LEP) was first identified and mapped to human chromosome 7q32.11994 (ob/ob) mice, which have a leptin mutation, triggering many comorbidities such as obesity, hyperinsulinemia, corticosterone excess, and infertility  were revised , 66–71.LEP described in 1997 by Montague et al. (1997)  . Two couIn 2002, a third leptin-deficient child (Ob3) from a consanguineous family of Pakistani ancestry was identified homozygous to ΔG133 by the same group . HoweverTwo years later, a fourth pediatric case (Ob4) homozygous for ΔG133 was reported . Like thLEP sequence of each case was determined, and seven were identified homozygous for the ΔG133 frameshift mutation, indicating this variant as a founder mutation in Central Punjab. One case was homozygous for a novel frameshift mutation in the exon 3 (p.Leu161fsX170), as a result of CT deletion in codon 161, shifting the stop codon position and disrupting leptin folding sites.A study conducted by Fatima et al. 2011) (11 (20) iLEP variants were identified in 10 (16.1%) unrelated probands from consanguineous families, of these, nine (14.5%) were homozygous carriers for the previously mentioned frameshift mutation (ΔG133). All nine carriers were leptin deficient, and three subjects exhibited hyperinsulinemia. Interestingly, eight out of nine families with ΔG133 frameshift mutation belonged to the Arain community, which has a tradition in consanguineous marriages and is part 1 of the larger subethnic populations of Punjab. Like in the earlier studies, it was concluded that ΔG133 may be a founder mutation. In another study by Saeed et al. (2014)  , two unrAnother work, also performed by Saeed et al. (2015) , extendeThe most recent cross-sectional study conducted by ElSaeed et al. (2020)  revealedDeletion mutations can affect a single nucleotide, large sequences, or whole chromosomes. The aforementioned study carried out with Pakistan infants by Fatima et al. 2011) (11 (20), A novel mutation caused by a 42-pb deletion in intron 1 was identified by Saeed et al. (2015)  in a malLEP human gene was identified in a 6-month-old boy, with consanguineous parents. The patient was severely obese with remarkable hyperphagia and low leptin serum level. Additionally, the patient presented with central hypothyroidism, elevated liver enzymes, dyslipidemia, and grade 2 hepatic steatosis. The electrophoresis revealed the lack of a fragment of 2,862 bp in the PCR product of the patient. This missing fragment corresponds to exons 2 and 3 of LEP. These findings indicate a homozygous deletion of the leptin coding sequences in an infant proband . Novel information about patients 14, 24, and 31 were also later reported . PatientLEP gene (p.Arg105Trp)  . The prog105Trp) .LEP variants, including two siblings homozygous to p.Arg105Trp. A third affected sibling died at the age of 3. The two alive probands were 1.5- and 10-year-old males and were the first Pakistani identified with this leptin variant   also desLEP p.Asn103Lys and wild type and concluded that the leptin mutant is secreted, however, not functional (A novel leptin missense in homozygosigotic mutation caused by a C-to-A substitution in the third base of codon 103 (p.Asn103Lys), located in the protein N-terminus region was described in two Egyptian siblings, a 3-year-old boy and a 7-year-old girl. Both had early-onset severe obesity, remarkable hyperphagia, hyperinsulinism, and low leptin serum levels. The probands were the result of consanguineous marriage, in which the parents were heterozygous variant carriers. The probands had a history of delayed development, but no other clinical features to suggest syndromic obesity . This vanctional .The presence of p.Asn103Lys leptin mutation was also investigated in the Pakistani population since Pakistan is the 9th country among 188 countries in obesity ranking. A case-control observational study enrolled 475 unrelated subjects, of which 250 were obese. The homozygous mutation was identified in a 10-year-old boy from a highly consanguineous family with several cases of obesity. The proband had the classical features observed in obesity caused by congenital leptin deficiency, with leptin level close to limit detection. The authors then hypothesized that this mutation, besides processing inactive leptin, also may be associated with low leptin serum levels .A 14-year-old girl from Austrian origin, with mild obesity, was identified with a homozygous transition in exon 3 (thymine to cytosine), resulting in leucine to serine exchange in codon 72 of leptin protein. She presented with undetectable leptin, and remarkable clinical history of rapid weight gain leading to obesity after normal birthweight, even after restricted caloric diet. She was the offspring of healthy and nonconsanguineous parents heterozygous to this mutation. The proband entered puberty but exhibited features of hypogonadotropic hypogonadism. She had hyperinsulinemia, increased transaminases, and dyslipidemia in addition to sympathetic system dysfunction observed under cold pressor test. No immune abnormalities were observed, with normal T-cell counts and activity. Function assays proved that Leu72Ser leptin variant is expressed but not secreted; however, in contrast to other cases with congenital leptin deficiency, this patient only showed mild obesity, probably due to a residual leptin activity or due to the patient’s daily environment, favorable to controlling energy intake since infancy .LEP coding region of 35 obese cases with BMI ≥32 kg/m² and controls with BMI <25 kg/m². The His118Leu mutation in exon 3 of LEP was described for the first time in one obese patient with a BMI of 46.0 kg/m². In addition to obesity, he presented with hypertension, metabolic syndrome, fatty liver, sleep apnea, gastric ulcer, and chronic superficial gastritis. The current mutation was accessed using the predicting bioinformatic tools PolyPhen2 and SIFT, which classified the mutation as “damaging.” Pedigrees and anthropometric and biochemical data regarding the patient were not available, and functional analyzes of this novel variant were not performed. Further studies to address these limitations are required   also ideA case report of an Indian infant at 10 months of age described classical features of congenital leptin deficiency, including low serum leptin concentrations. She was a child of healthy, nonobese, consanguineous parents. Genetic analysis identified an amino acid change of asparagine to aspartic acid in position 100 (p.Asp100Asn) of the protein. Prediction effect analysis classified this novel variant as “probably pathogenic” and “pathogenic” by PolyPhen-2 and by SIFT, Log ratio test, and MutationTaster, respectively .LEP identified both sisters homozygous for a novel missense mutation in codon 117, a cysteine to phenylalanine substitution. This was the first leptin variant reported on the American Continent. The older sister was 24 years old and had primary amenorrhea. At 16 years old, her BMI reached 53 kg/m², and bariatric surgery was performed, resulting in a weight loss of 20 kg, which was not sustained. She presented with increased weight gain in the following 5 years, even under restricted sugar and fat diet and 60 min of walk daily. The younger sister was 21 years old, with primary amenorrhea in addition to hypertriglyceridemia, insulin resistance, and acanthosis nigricans. Both patients exhibited breasts and genitals at Tanner Stage V, and undetectable leptin serum levels . Her parents were heterozygous for this mutation . History of respiratory tract infection was reported to the male proband. It was also reported that two younger siblings, also with severe obesity and with recurrent respiratory infection, had died. The parents were heterozygous carriers .Studies with administration of recombinant leptin in mice began in 1995 and showed that its use ameliorated their obesity through reduction of food intake and higher energy expenditure –86. The The replacement therapy with metreleptin in the Ob1 patient, one of the first individuals identified homozygous to ΔG133, was applied . The synad libitum test meal. Before treatment, Ob2 and Ob3 patients exhibited CD4+ lymphopenia and deficient lymphocyte activity; however, the therapy with the synthetic leptin addressed these issues, and both patients showed normal immunophenotype.Later in 2002, a longer study of leptin replacement therapy, ranging from 10 to 50 months, included the Ob1, Ob2, and Ob3 patients . As obseThe fourth case carrying the ΔG133 variant (Ob4) also was benefited from leptin replacement therapy. The patient showed some episodes of weight gain, which was contoured with leptin dose adequacy. Ob4 had a decrease of total fat mass of 15.9 kg with 48 months of therapy, and BMI lowered to 24.2 kg/m². Biochemical parameters and immune function were improved, with attention to the amelioration of perineal dermatitis, asthma symptoms, and fewer occurrences of urinary tract infections. These results are in line with the observations from all previous cases homozygous for ΔG133 after r-metHu-Leptin therapy. The authors concluded that the abnormal thyroid function of Ob4 was completely normalized after leptin replacement and T4 therapy withdrawal .Three adult probands from the Turkish family identified with the p.Arg105Trp mutation , 24 wentad libitum test meal. To examine the brain activity of the probands before and after therapy, functional magnetic resonance imaging was used. The patients were shown images of food and nonfood during fasting and after feeding and were asked to pick the images they liked. Leptin replacement was associated with a decreased “liking” rate from patients who were shown images of food. Before the leptin treatment, the accumbens-caudate was activated either during the starving state and feeding state. After the therapy, accumbens-caudate activation was found to be correlated only with the starving state, suggesting a role for leptin in food reward perception and satiety signaling.The effects of leptin replacement on eating behavior were accessed by Farooqi et al. (2007)  in two lIn light of the findings above, Paz-Filho et al. (2008)  aimed toFinally, Wabitsch et al. (2015b)  reportedTaken together, these studies showed that the use of recombinant leptin improves the clinical spectrum of leptin-deficient patients. The use of the exogenous leptin results in quality-of-life improvements.LEP gene discovery in 1994 to the application of exogenous leptin to overcome congenital leptin deficiency, much progress has obviously been made. This adipokine has a unique neuroendocrine role regulating energy expenditure, food consumption, and the hypothalamic-pituitary-gonadal axis. Homozygous LEP autosomal recessive mutations are correlated to the expression of disease phenotypes including hyperphagia, hyperinsulinemia, immune system dysfunction, and infertility. Since the description of the first monogenic variant, a total of 17 mutations in 67 cases were reported in the literature, with the majority reported from highly consanguineous families , National Council for Scientific and Technological Development (CNPq), Carlos Chagas Filho Foundation for Research Support in the State of Rio de Janeiro (FAPERJ), and Coordination of Superior Level Staff Improvement (CAPES). The funding source had no involvement in study design; in the collection, analysis, and interpretation of data; in the writing of the report; and in the decision to submit the article for publication.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.


The growing interest in BCL2 inhibitors for the treatment of multiple myeloma (MM) has led to the need for biomarkers that are able to predict patient’s sensitivity to the drug. The presence of the chromosomal translocation t in MM is mainly associated with sensitivity to venetoclax and good prognosis. The incidence of t largely increases in primary Plasma Cell Leukemia (pPCL) in association with an unfavorable outcome. Currently, data concerning pPCL sensitivity to venetoclax are virtually absent. In this context, we investigated the transcriptome of MM and pPCL with t, evidencing that the two clinical entities are likely responsive to venetoclax based on different molecular programs, thus prompting further studies to elucidate better novel potential predictive biomarkers.Mechanisms underlying the pathophysiology of primary Plasma Cell Leukemia (pPCL) and intramedullary multiple myeloma (MM) need to be further elucidated, being potentially relevant for improving therapeutic approaches. In such a context, the MM and pPCL subgroups characterized by t deserve a focused investigation, as the presence of the translocation is mainly associated with sensitivity to venetoclax. Herein, we investigated a proprietary cohort of MM and pPCL patients, focusing on the transcriptional signature of samples carrying t, whose incidence increases in pPCL in association with an unfavorable outcome. In addition, we evaluated the expression levels of the BCL2-gene family members and of a panel of B-cell genes recently reported to be associated with sensitivity to venetoclax in MM. Moreover, transcriptional analysis of lncRNAs in the two clinical settings led to the identification of several differentially expressed transcripts, among which the SNGH6 deregulated lncRNA might be relevant in the pathogenesis and prognosis of pPCL with t. Overall, our data suggest that MMs and pPCLs with t might be responsive to venetoclax based on different molecular programs, prompting further studies to elucidate better novel potential predictive biomarkers. Multiple myeloma (MM) is a malignant proliferation of antibody-secreting bone-marrow plasma cells (PCs) that is characterized by a highly heterogeneous genetic background with structural chromosomal aberrations and specific gene mutations . MM has Several studies investigating the gene mutation patterns  and diffIn the present study, we investigated the transcriptomic profiles of MM and pPCL samples carrying t in a proprietary gene expression profile dataset (GSE116294). In addition, given the increasing importance of BCL2 inhibitors in the treatment of MM, both MMs and pPCLs stratified for the presence of t were evaluated for the expression levels of the BCL2 gene family and of a panel of B-cell genes recently reported as associated with venetoclax sensitivity in MM . Our anaFull details of multi-omics data in CoMMpass study and statistical analysis are provided in the Supplementary Methods.We investigated, by gene expression array, a proprietary cohort of 50 MM patients and 15 primary PCLs (pPCL) that was purified and characterized as previously described . This co® Human Gene 2.0 ST arrays , thus obtaining the expression level of 18,642 protein coding transcripts and 10,138 unique lncRNAs, as previously described [https://github.com/MikeJSeo/SAM, accessed on 12 May 2021), as previously described [p-value < 0.05. All the data are available in the NCBI Gene Expression Omnibus database  and are accessible under accession #GSE116294 [Fifty MMs, 15 PCLs, and 4 N samples were profiled on GeneChipescribed . The lisescribed . Gene seSE116294 .We performed qRT-PCR as previously described . PrimersAt first, we analyzed the transcriptome of MM and pPCL carrying the t translocation by investigating the expression profiling of a cohort of 50 newly diagnosed MM patients representative of the major molecular types of the disease, 15 pPCLs, and four normal controls .® Human Gene 2.0 ST array, which is able to detect 35.458 transcripts, including 18.642 protein coding (53%) and 10.138 lncRNAs (29%), upon annotation on unambiguous entries in GENCODE encyclopedia (V25) [We took advantage of the GeneChipia (V25) . HierarcThe 2402 most variable transcripts driving the clustering are mainly enriched in protein coding genes (66%) but also include lncRNAs (15%), as well as a heterogeneous group of transcripts among which immunoglobulin genes, small nucleolar RNAs, miRNAs, and pseudogenes. Based on these considerations, we focused our further studies on protein-coding genes and lncRNAs.To identify the genes that specifically distinguished the two clinical entities in the context of t, we performed a supervised analysis between the seven pPCL and the 12 MM cases carrying the translocation. A comprehensive list of 2416 differentially expressed (DE) protein-coding genes  was obtained at a low stringency cutoff (FDR < 10%) . In the In addition, to define those molecular pathways differently modulated in relation to MM and pPCL with t, a Gene Set Enrichment Analysis (GSEA) was performed on the list of 628 DE protein-coding genes that were ranked based on fold-change values . RemarkaIn detail, among the significantly enriched gene sets , we evidGiven the increasing importance of BCL2 inhibitors in the treatment of MM and the enrichment of the responders in patients with t, we investigated the expression levels of all the BCL2 gene family members in both MM and pPCL samples stratified for the presence or absence of the t, and normal controls .Concerning the anti-apoptotic proteins, a significant overexpression of BCL2 was found in MM patients with t compared to the normal controls, MM cases not carrying the translocation and pPCL samples, irrespective of the presence of the t. On the other hand, we observed a significant downregulation of BCL2L1 (known to encode for BCL-XL) in all pathological samples compared to normal controls, and in particular in patients carrying t, markedly in pPCL. Therefore, when considering the BCL2/BCL2L1 ratio, known as an essential response predictor to venetoclax ,27, it wAmong genes encoding sensors for cellular stress and DNA damage, higher PMAIP1 expression levels were associated with the presence of t in both MM and pPCL, although PMAIP1 overexpression in t MMs did not reach significance compared to normal controls. Conversely, a lower BMF expression was significantly associated with the absence of t only in the MM group.Regarding apoptosis activator proteins, higher BCL2L11 expression levels were associated with t in MM. Finally, focusing on apoptosis effector proteins, we found that, in both MMs and pPCLs, BAK1 overexpression was significantly associated with the absence of t, being also significant if compared to normal controls; in addition, the median lowest BAX expression was observed in t MMs, although not reaching significance when compared to N controls.The expression levels of BCL2, BCL2L1, and MCL1 were validated by qRT-PCR in 11 MM and five pPCL samples profiled by gene expression arrays for whom RNA material was available  CoMMpass dataset including MM samples characterized for genome abnormalities, transcriptome, and clinical outcome, we confirmed that BCL2, BMF, PMAIP1, and BCL2L11 expression levels were higher in MMs with t than in MMs without t. In addition, we validated the lower expression levels of BCL2L1, BAK1, and BAX in MMs carrying t compared to the other MM group. Moreover, similarly to our proprietary database, in the CoMMpass cohort no differences in the expression of MCL1, BCL2A1, BBC3, and BAD were detectable in MMs with or without the t. Concerning BCL2L2, HRK, and BID, whose expression levels did not reach any relevant difference between the two MM groups in our database, probably due to the limited number of patients, in the CoMMpass cohort their expression was significantly higher in MMs with t .Recent data described a gene expression signature enriched for B-cell-associated genes in MM cell lines and MM patients sensitive to venetoclax . Given tAs for BCL2 family genes, MS4A1, CD79A, SORT1, and BEND5 expression levels were validated by qRT-PCR in 16 samples profiled by gene expression arrays for whom RNA material was available  were confirmed in the CoMMpass dataset. In detail, VPREB3, MS4A1, CD79A, STAT5A, SORT1, BEND5, and REL genes were all significantly upregulated in MMs with t compared to the other MM subgroup. Moreover, BATF expression level was significantly lower in the MM group with t, whereas no differences in the expression of the RASGRP2 gene were detectable between the two MM groups in CoMMpass cohort. Concerning CXCR5, PIK3AP1, and IL4R, for which differences in expression levels between the two groups in our database did not reach significance, probably due to the limited number of patients, in the CoMMpass cohort, their expression was significantly higher in MMs with t .Next, we investigated lncRNA transcriptional patterns specifically differentiating pPCL and MM cases carrying t. In detail, we performed a supervised analysis between the 7 pPCLs and the 12 MM cases carrying such a translocation, detecting 38 DE lncRNAs . As for protein-coding genes, we excluded from further analyses the five lncRNAs in common with the analysis comparing all 15 pPCL and 50 MM cases . Therefoq-value < 0.001) was observed for 26 lncRNA-gene pairs; among them, we identified 10 lncRNA-coding genes couples  and Progression-Free Survival (PFS), freely accessible from MMRF Study, were analyzed in 767 MM cases with available RNA-seq data. In detail, for each lncRNA we defined two MM patient groups based on their median lncRNA expression level see . Kaplan–The clinical impact of these five lncRNAs was further investigated by Cox regression univariate analysis. For three lncRNAs, their lower expression level was associated with a significantly higher risk in PFS, and for one of them (Linc00886) also in OS . Conversp = 0.037). Concerning the copy number alterations (CNAs) commonly found in MM, 1q gain/amp occurred significantly more frequently in patients with low SNHG6 expression levels (p = 0.0074). Finally, the SNHG6 expression level was not correlated with the presence of mutations affecting K-RAS, N-RAS, BRAF, FAM46C, TP53, and DIS3 genes , whose incidence increases in pPCL in association with an unfavorable outcome. These selection criteria allowed us to investigate possible molecular mechanisms other than those associated with this chromosomal abnormality that could better characterize the differences between these two clinical and pathological forms associated with the t. We found that the two clinical entities are distinguishable at the transcriptomic level despite sharing the same t genetic background. In particular, we defined the specific signatures of coding genes and lncRNAs that discriminate MM and pPCL samples with t by refining from the lists those transcripts that significantly differentiate the two clinical settings in general.Our data showed that the pPCL group with t was particularly enriched in genes involved in IL2-STAT5 signaling , which iThe interest in BCL2 inhibitors in MM treatment is continuously increasing, especially for venetoclax that is capable of inducing apoptosis in cells dependent upon BCL2 for survival. Most myelomas are MCL1 dependent; however, a subset of MM enriched for t is co-dependent on BCL2 and thus sensitive to venetoclax. Indeed, in numerous pre-clinical and clinical studies venetoclax has been reported to be consistently more effective in MM harboring the t translocation ,33,34. HIn detail, we found that MM and pPCL cases with t showed a distinctive expression pattern of the BCL2 family genes , althougBesides the distinct expression pattern of BCL2 family genes, MM and pPCL cases with t largely differed for the expression of B-cell genes associated with venetoclax sensitive MM . Indeed,Overall, these data suggest that MMs and pPCLs may be sensitive to venetoclax based on different molecular programs, implying that specific biomarkers should be investigated in the two clinical entities as the best predictor of venetoclax sensitivity.Concerning lncRNAs, our data indicated that MMs and pPCLs with t scarcely differed from each other based on lncRNA expression. However, some of the lncRNAs found in the signature are suggestive for pathogenic implication in pPCL and warrant further investigation. Indeed, AC107884.1, whose expression levels in pPCLs are significantly lower than in MM, is highly correlated with the OLFML1 gene , whose aTranscriptomic analyses of MMs and pPCLs with t indicated that the two clinical entities might be responsive to venetoclax based on different molecular programs, prompting further study to identify biomarkers of venetoclax sensitivity specific for MM or pPCL. Our results also suggested that SNHG6 deregulation might be relevant in the pathogenesis and prognosis of pPCLs with t.


Saccharomyces cerevisiae, Saccharomyces pastorianus and Saccharomyces pastorianus residual biomass on natural polymers  and to evaluate the biosorptive potential for removal of pharmaceuticals and dyes from water. Six types of biosorbents were synthesized and characterized by Scanning Electron Microscopy and Fourier Transform Infrared Spectroscopy techniques and their biosorptive capacities for three drugs  and two dyes (orange II and indigo carmine) were evaluated. The obtained results show that the removal efficiency depends on the polymer type used for the immobilization. In case of alginate the removal efficiency is between 40.05% and 96.41% for drugs and between 27.83% and 58.29% for dyes, while in the case of chitosan it is between 40.83% and 77.92% for drugs and between 17.17% and 44.77% for dyes. In general, the synthesized biosorbents proved to be promising for the removal of drugs and dyes from aqueous solutions.Pharmaceuticals and dyes are a very important part of the nonbiodegradable or hard biodegradable substances present in wastewater. Microorganisms are already known to be effective biosorbents, but the use of free microbial cells involves difficulties in their separation from effluents and limits their application in wastewater treatment. Thus, this study aimed to develop biosorbents by immobilizing  In the last few decades, due to the rapid industrialization and increasing of the world population, the ecosystems are seriously threatened due to the uncontrolled discharge of a large number of toxic organic or inorganic pollutants ,2,3,4. AMost of these inorganic and organic pollutants have accumulated in the environment due to their stability and difficult degradation and are detected in different environmental matrices: surface water, groundwater, soil and drinking water ,3,8,9.The World Health Organization states that water is an essential element for sustaining life, so water supply must be adequate, safe and accessible to all. Harmful effects on ecosystems and health hazards due to organic and inorganic pollutants present in water have been established over time, making it necessary to apply ever-higher standards for the detection and treatment of pollutants .To remove pollutants from water, several physicochemical and biological processes have been developed such as adsorption, biosorption, coagulation, sedimentation, filtration, ion exchange, membrane technologies and biological treatments ,12. SomePharmaceuticals are a very important part of the nonbiodegradable or hard biodegradable substances present in wastewater and effluents from treatment plants. The annual production of pharmaceutical compounds has been estimated at thousands of tons worldwide . PharmacThe presence of these pollutants in natural water poses a significant risk and some of them have already been declared as priority substances in water protection policies. Therefore, the development of innovative water treatment technologies is necessary to ensure water quality .Lately, research has focused on removing persistent organic pollutants from water using as methods: adsorption/biosorption or advanced oxidation ,18,19,20This has been and remains a promising biotechnology for the removal of pollutants from aqueous solutions due to its simplicity, efficiency and to the availability of biomass and residual biomass. The biosorptive capacities of different types of biomasses in relation to the removal of different types of pollutants have been reported in numerous scientific papers ,24,25,26Biosorption as a process is based on a variety of mechanisms including: adsorption, absorption, ion exchange, surface complexation and precipitation.From the first research on the pollutants biosorption processes, the studies focused on obtaining efficient, viable, economical and easy-to-apply in wastewater treatment, biosorbents.The literature presents numerous materials of biological origin that have been investigated to obtain biosorbents that include microbial biomass , microbial residual biomass (resulting from fermentation processes), agricultural waste, food industry waste and other materials ,23,27,28Both microorganisms and microbial residual biomass have been noted for good biosorptive capacity related to most pollutants tested. Still, they have a number of drawbacks, including the fact that free microbial cells are small in size and low in density, which involves difficulties in their separation from effluents and obviously, an increase in the cost of the biosorption process, which limits their application in wastewater treatment ,23.In this sense, the immobilization and encapsulation of microorganisms and residual biomass would be an alternative for obtaining biosorbents.Different immobilization techniques and different supports were tested, among which we can mention polysulfone, polyvinyl alcohol, polyacrylamide, polyurethane, silica gel, carboxymethyl cellulose, brewer’s spent grain, sodium alginate, chitosan ,10,23,24Natural polymers such as chitosan and alginate have been used successfully in the processes of immobilization/encapsulation of enzymes and pharmaceuticals ,30,31. SChitosan has been demonstrated that it can be used in adsorption processes because it is an inert, biocompatible, biodegradable and nontoxic material ,32,41.Alginate is also an alternative for immobilization due to its good chemical stability, nontoxicity, incorporation efficiency and low cost ,7,24,26.Saccharomyces cerevisiae is one of the most researched species in biosorption processes. Both Saccharomyces cerevisiae and Saccharomyces pastorianus are used in brewing and are available in large quantities.In this context, this paper focuses on obtaining eco-friendly biosorbents that can be used in the treatment of effluents containing persistent organic pollutants.Saccharomyces cerevisiae, Saccharomyces pastorianus and Saccharomyces pastorianus residual biomass on natural polymers  and to evaluate the biosorptive potential for removal of pharmaceuticals and dyes from water.This study aimed to develop biosorbents by immobilizing Six types of biosorbents were synthesized and characterized by scanning electron microscopy (SEM) and Fourier transform infrared spectroscopy (FTIR) techniques and their biosorptive capacities for three drugs , rifampicin (RIF), ethacridine lactate (EL)) and two dyes (orange II (O II) and indigo carmine (IC)) were evaluated.-CPX is an antibiotic which is considered an emerging compound with a risk factor greater than 1 in a number of countries ;-RIF is an antibiotic which is a persistent compound in water matrices with high potential to induce antimicrobial resistance in bacterial cells, leading to mutagenic and carcinogenic effects ;-EL is an antiseptic and local analgesic used in large quantities worldwide which can be found in effluents of wastewater treatment plants. Therefore it represents a potential risk for aquatic and other environments .The pharmaceutical compounds were chosen as target molecules taking into account the following considerations:As for the selected dyes, both IC and O II are used in various industrial sectors , discharged in considerable amounts in water and recognized as recalcitrant compounds in water policies, difficult to remove by conventional water treatment ,44.Saccharomyces pastorianus residual biomass immobilized on natural polymers related to removal of drugs and dyes from aqueous solutions.Our literature survey revealed that RIF, EL, IC and O II are less studied from the point of view of their removal by adsorptive/ biosorptive processes. Therefore, the present study aims to provide new insights into application of these types of biosorbents in the removal of drugs and dyes from water. To the best of our knowledge, this is the first study to evaluate the biosorptive capacity of All reagents implied in the experimental step were of analytical grade and were used without further purification. Sodium alginate and chitosan were bought from Carl Roth, Karlsruhe, Germany; sodium hydroxide and calcium chloride were delivered by Chempur, Piekary Śląskie, Poland; hydrochloride acid, sodium chloride, acetic acid, di-sodium hydrogen phosphate, potassium di-hydrogen phosphate, bentonite and ethanol were purchased from Chemical Company, Iași, Romania; cephalexin was from Cayman Chemical, Tallinn, Estonia; orange II and indigo carmine green were procured from Merck, Darmstadt, Germany.Saccharomyces cerevisiae and Saccharomyces pastorianus and residual biomass of Saccharomyces pastorianus were provided by two local companies .Dried yeast strains of Stock solutions of drugs and dyes with concentrations of 500 mg/L were prepared by dissolving the reagents in distilled water and kept at 4 °C in closed recipients.For the calibration curves, adequate volumes of stock solutions were placed in series of 10 mL volumetric flasks. Concentrations ranges were between 1 mg/L and 30 mg/L for CPX; 1 mg/L and 150 mg/L for EL; 1 mg/L and 50 mg/L for RIF; 1 mg/L and 15 mg/L for O II and between 1 mg/L and 20 mg/L for IC.Samples absorbance were acquired with the help of a UV1280 spectrophotometer  at 260 nm for CPX, 431 nm for EL, 475 nm for RIF, 485 nm for O II and 610 nm for IC. Calibration graphs (absorbance vs. concentration) served to recover linear regression equations.The solutions’ pH levels were adjusted with NaOH (0.1 M) or HCl (0.1 M).Saccharomyces cerevisiae/calcium alginate (SC-A-5%) and Saccharomyces pastorianus/calcium alginate (SP-A-5%) beads preparation, aliquots of sodium alginate were placed in laboratory beakers containing hot water and mixed ) until complete dissolution. Dried yeasts strains were added to final concentrations of 5% on dry weight (d.w.) basis. The mixing process was continued until homogeneous suspensions were obtained. The resulting combinations were suspended in 2% calcium chloride solutions and kept at 4 °C.For Saccharomyces cerevisiae/chitosan (SC-C-2.5%) and Saccharomyces pastorianus/chitosan (SP-C-2.5%) beads were synthesized according to the method presented by Kaushal et al. [l et al.  with somSaccharomyces pastorianus residual microbial biomass, the procedure used by de Rossi et al. [For the biosorbents prepared with i et al.  was consA Quanta 200 3D  scanning electron microscope (SEM) with energy-dispersive X-ray system and a large field detector (LFD) was used for the surface morphology and elemental composition analyses of the synthesized biosorbents. The dried samples  were fixed to stubs with double adhesive carbon discs. Secondary electron mode (SE) in low vacuum was applied at an accelerating voltage of 20 kV, a working distance of 15 mm and a spot size of 5. The magnification range was between 1 mm and 10 µm.−1 and 400 cm−1 (32 scans coadded) with a resolution of 4 cm−1 on a Nicolet iS50 FT-IR spectrometer  coupled with a built-in ATR accessory, DTGS detector and a KBr beam splitter. ATR ethanol cleaning was performed after each spectrum acquisition. The background spectrum reference with air was recorded and compared with the anterior one.Fourier transform infrared (FT-IR) spectra were registered between 4000 cmi) settled between 2 and 12 with HCl (0.1 M) or NaOH (0.1 M) and measured with a portable pH meter  were stirred on magnetic plates with 0.5 g of biosorbent for 24 h, at room temperature. Then, the pH values (pHf) were measured again. The pHpzc were retrieved from curves pHf = f(pHi).For the determination of biosorbents points of zero charge, the following modus operandi was applied. Volumes of 25 mL of 0.1 M NaCl solutions with initial pH values  and dyes  solutions were put in contact with 1 g of biosorbents beads for 12 h, at ambient temperature.The residual concentrations were calculated by reading the samples absorbance at dedicated wavelength against the calibration curves.R, %) and the biosorption capacity  were determined with the Equations (1) and (2).C0 and fC are the initial and final concentrations (mg/L); V is the sample volume (L) and m is the biosorbent amount (g).The removal efficiency  and 1576 cm−1 to C=O stretching (amide I) from chitosan but also specific for yeast cell walls constituents; 1363 cm−1 to CH3 vibration [For SC-C-2.5% and SP-C-2.5% adsorption bands can be attributed as follows: 2920 cmibration ,61.−1 and at 1238 cm−1 stretching for amide III (proteins) and for asymmetric and symmetric PO2− (phosphorylated proteins and phospholipids) were detected and were caused by the increased amount of yeast.In SPRMB-A-5% and SPRMB-A-9% at 1318 cmpzc), defined as the pH value at which the biosorbents’ surface charge is null, i.e., the charge of the positive surface sites is the same as that of the negative ones, was determined with the drift method with sodium chloride as electrolyte. As shown in f with the augmentation of NaCl pHi, with a plateau when the pHi is between 4 and 10 and with a new increase step after pHi 10. The same evolution was mentioned by Kragovic et al. [Point of zero charge . Similar values were encountered when Bacillus subtilis bacterial biomass was used without treatment to remove CPX from aqueous systems [For the antibiotic cephalexin, the best biosorption capacities were recorded for the biosorbents prepared on chitosan (22.78 mg/g for SC-C-2.5% and 28.42 mg/g for SP-C-2.5%) and the lowest ones for those prepared by the immobilization of residual microbial biomass of  systems ,64. As r systems , the retIn the case of ethacridine lactate, the removal efficiencies were between 54.28% for SPRMB-A-5% and 96.40% for SC-A-5%. At present, very little information is available for the elimination of this molecule by adsorption. One of the publications dedicated to such study  reveals Another of the tested molecules was the rifampicin, an antibiotic with a common structure consisting of a naphtohydroquinone chromophore spanned by an aliphatic ansa chain . The recRegarding the dyes’ biosorption, from In our tested conditions, the prepared biosorbents gave reasonable results in terms of the ability of removing drugs and dyes from aqueous solutions. However, it should be mentioned that in optimized settings, better uptake efficiencies could be expected.The synthesized biosorbents exhibited similar characteristics, which proves that the techniques used for their synthesis are efficient.The immobilization methods of microorganisms and residual biomass in the polymer matrix of alginate and chitosan were shown to be effective, the cells’ immobilization being confirmed by SEM and FT-IR analyzes.PZC values depend on the polymer type used for the immobilization and not the type of biomass that was used.The points of zero charge were identified at 6.9 for SC-A-5%, 6.6 for SP-A-5%, 6.9 for SPRMB-A-5% and 6.8 for SPRBM-A-9%, 8.3 for SC-C-2.5% and 8.8 for SP-C-2%. The pHThe obtained results show that the removal efficiency depends on the type of biosorbent and the concentration of the pollutant. In the case of using the alginate for immobilization the removal efficiency is between 40.05% and 96.41% for drugs and between 27.83% and 58.29% for dyes, while in the case of chitosan it is between 40.83% and 77.92% for drugs and between 17.17% and 44.77% for dyes. The biosorption capacities obtained in the presented test conditions vary between 4.79 mg/g and 85.42 mg/g for drugs and between 4.34 mg/g and 26.87 mg/g for dyes. This indicates a good biosorption capacity if we take into account the amounts of pollutants present in different water matrices.The mentioned results reveal that all the synthesized biosorbents can be useful for the removal of pharmaceuticals and dyes from aqueous solutions. Further, by using the immobilization technique on natural polymers, first of all, the disadvantages of using free cells are eliminated and secondly, eco-friendly materials are obtained.Finally, additional work is needed to study the effect of process parameters  in order to optimize biosorption capacity.


Background: The findings of associations between prenatal air pollution exposure and hypospadias risk in offspring are inconsistent. No systematic review or meta-analysis has yet summarized the present knowledge on the aforementioned topic.Methods: Relevant manuscripts were identified by searching PubMed and Web of Science databases through January 31, 2020. Summary odds ratios (ORs) with 95% confidence intervals (CIs) in meta-analyses were estimated based on a random effects model. Publication bias was evaluated by funnel plots, Begg’s test, and Egger’s test.2.5 exposure in the first trimester was related to increased risk of hypospadias .Results: The search identified 3,032 relevant studies. Sixteen studies cumulatively involving 21,701 hypospadias cases and 1,465,364 participants were included. All of these studies were classified as having a low risk of bias. We classified pollutants as nitrogen oxides, particulate matter (PM), ozone, and other exposures. The exposure window to pollutants varied from three months before conception to seven days after delivery. In the meta-analyses, only PM2.5 exposure on hypospadias risk. Improvements in the areas of study design, exposure assessment, and specific exposure window are needed to advance this field.Conclusion: We found evidence for an effect of PM Increasdiseases , and cardiseases . Notablydiseases –8. Amongdiseases , 10. Prediseases –14.2.5 and heavy metal hazardous air pollutants increased hypospadias risk [2, PM10, PM2.5, and heavy metal hazardous air pollutants (HMHAPs) in early pregnancy [Hypospadias, which occurs between 8 to 16 weeks of gestation , is one ias risk , 17–21. regnancy –22. The regnancy –14, mainThe detailed process of study selection is illustrated in The key characteristics of the included studies are reported in The risk of bias of included studies was evaluated according to OHAT criteria. Overall, low risk was assessed for most of the studies in this systematic review, especially for the selection bias, confounding bias, and performance bias . It was Due to the lack of basic information provided for the control group, two studies , 25 wereIn general, we observed consistent results across the risk assessments for bias in different studies. The analogy in study design explains the consistency of the bias rating risks allocated across the studies. Our literature search revealed 16 studies considered of sufficient quality to prove the conclusions of our systematic review.X [2 [2 exposure [X, NO2, and NO exposures in the 3 months post conception were mainly positively associated with hypospadias though there was no statistical significance. Furthermore, Huang et al. [X as well as NO2 exposure and hypospadias risk during the 3 months before pregnancy and other months after pregnancy. No statistically significant results were found. However, Salavati et al. [X and NO2 during the periconceptional period.Three, five, and one studies were carried out to investigate the relationship between prenatal NOX , 17, 20,X [2 , 31, andX [2  exposureexposure , 31, twoexposure . In genei et al.  found a 10 [2.5 [2.5–10 [10 and PM2.5 exposure during 3 months after pregnancy, three, three, and one studies reported an insignificant positive association [2.5–10 exposure and hypospadias risk.A total of 10 studies (three cohort and seven case–control studies) investigated the connection between PM exposure and hypospadias risk in seven countries, including the USA, Spain, Italy, the Netherlands, Britain, China, and Israel. Seven, eight, and two studies focused on prenatal exposure to PM10 , 29, 31,10 [2.5 , 28, 31, [2.5–10 , respectociation , 18, 20,ociation –29, and ociation . Of noteociation  and Huan3 increments in PM2.5 by the first trimester period [2.5 during the first trimester was associated with higher odds of hypospadias  with moderate heterogeneity   . Visual = 0.039)  and the 3 exposure and hypospadias risk [3 exposure with the risk of hypospadias [3 during the first trimester were not significantly associated with hypospadias risk   . In addi3 exposure and hypospadias risk in the first month after conception. Of note, Huang et al. [3 and hypospadias risk, but no statistically significant result was found.However, Huang et al.  observedg et al.  also focg et al. , 27 inve2, CO, landfill sites, garbage wastes, hormonally active hazardous air pollutants, and HMHAPs in seven countries, including Belgium, Denmark, France, Italy, Britain, the USA, and Israel. Six studies focused on the exposure window in the prenatal period [Seven case control studies and two cohort studies were included , 30, 31.l period –26, 30 al period , 27, 31.l period  observedl period  found a 2.5 exposure during the first trimester was related to statistically significant increases in the risk of hypospadias, but no statistically significant increase in the risk of hypospadias in relation to continuous O3 exposure during the first trimester. In the present study, we did not perform further meta-analyses to assess hypospadias with other air pollutants or exposure windows because of the small number of studies presented.As the first systematic review summarizing the association between prenatal exposure to air pollution and hypospadias risk, our study added depth and clarity to the current evidence exploring the aforementioned topic. Meta-analysis suggested that continuous PMA few established biological processes that occur in the first trimester are related to the development of the urogenital system. Any interference happening in the pivotal window of internal and external urogenital development could become a cause of hypospadias . RecentlOur systematic review had several strengths. Compared to the limited sample size of individual studies, the present review comprehensively included all published observational studies (n = 16), which had relatively large sample sizes. The OHAT risk of bias rating tool was applied to systematically identify underlying risk of bias associated with multiple fields of studies included, while the NOS was used to assess the quality of the studies. Of note, the risk of bias in the included studies was low.10. Moreover, large well-designed cohort analyses are warranted in the future to explore whether there is a crucial association between air pollution and hypospadias risk.Although we conducted a meta-analysis of some exposures, several challenges should be taken into consideration. First, published studies included in this meta-analysis were too few to carry out a comprehensive analysis. Although studies were each of reasonable quality, high heterogeneity was generated by the different study designs, geographical locations, exposure windows and assessments, and cofounding adjustments. This may have affected the accuracy of the quantifiable findings in the meta-analysis. For instance, Vinceti et al.  reportedSecond, we could not exclude potential biases due to the misclassification of exposure and outcome. Using a fixed monitor to specify individual-level exposure within a specific radius around the monitor, including land-use regression models, has been widely accepted . HoweverThird, the criteria for considering eligible studies for systematic review should be based on a more rigorous PECOS structure. For example, as for exposure, the women in the first trimester of pregnancy were combined as a whole for the meta-analysis because of the small number of articles included. It is well-known that embryonic growth and development occurs in the first trimester of pregnancy, which is the critical time when most teratogenic exposures result in birth defects, including hypospadias. Of note, if the high concentration of certain pollutants or their metabolites accumulate for a long time in the pre-pregnancy period, it may cause a more obvious risk of congenital abnormalities . InteresFourth, besides the variations in method of measurement and exposure window, the strategies of analyses and forms of results reporting have been different among these studies. Based on the distribution of exposure in each study, air pollution was divided into dichotomy, trisection, and quartering. However, because of the different populations and exposure levels of each study, these aforementioned categories of exposure made it surely complicated to explain and compare the results. Dose-response analysis using a continuous exposure assessment model may provide more informative and nonbiased results, and more accurately reflect the association between ambient air pollution and birth defects than categorical variables, with higher levels of evidence.Finally, the importance of subgroup data should be recognized and, if possible, future studies should include subgroup analyses of the most relevant socio-demographic and clinical characteristics. Because all individual studies were observational in nature and multivariable analyses were carried out in the primary studies, the chance of residual confounding bias from unscanned variables cannot be excluded. More subgroup analyses stratified by these important potential confounders should be performed in future studies.3 increments in PM2.5 in the first trimester increased the risk of hypospadias. However, we could not determine the relationship between other ambient air pollutants and hypospadias risk. Further studies that focus on other exposures , as well as a more accurate assessment of exposures, better case ascertainments, and adjustments for a large number of potentially confounding effects, are needed to provide more evidence toward this topic.In summary, the present meta-analysis indicated that 10 μg/mThis systematic review and meta-analysis were based on the Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines  and the We implemented a compositive search in PubMed and Web of Science databases to determine all potentially correlative articles from inception through January 31, 2020. The following critical keywords were applied in the literature search: (air pollution) OR (traffic pollution) OR (outdoor pollution) OR (outdoor air pollution) OR (particulate matter) OR (nitrogen dioxide) OR (sulfur dioxide) OR (sulphur dioxide) OR (ozone) OR (carbon monoxide)) AND ((hypospadias) OR (hypospadia) OR (isolated hypospadias) OR (birth defects) OR  OR ). In addition, the bibliography of all included articles and related reviews and meta-analyses were further reviewed to determine whether there are other eligible articles. Searches were limited to English language articles. There is no geographical restriction.Two independent authors (ZX and SZ) deleted duplicates, filtered titles and abstracts for relevance, and determined records as included, excluded, or inconclusive. In case of uncertainty, the two authors obtained the full-text to determine eligibility. Any difference was settled through discussion and negotiation.Eligible studies were determined on the basis of the following PECOS  criteria: i) population: our study drew participants from pregnant women and their newborns; ii) exposure: the primary exposure of interest was air pollution on pregnant women; iii) comparison/comparator: the concentration of air pollutants to which the pregnant women were exposed; iv) outcome: the primary outcome of interest was the risk of hypospadias in offspring; the outcomes of other birth defects were not included; and v) study type. We targeted observational study designs ; studies without original data (review articles) were not included.Data extraction was carried out by SZ and confirmed independently by another author (ZX). The following data were from eligible studies: first author’s family name, publication year, study location, time period, study design, sample size, exposure characteristic , and main findings of the results.Two authors (ZX and SZ) independently assessed the quality of the observational studies using the Newcastle-Ottawa Scale (NOS) . The scaThe Office of Health Assessment and Translation (OHAT) risk of bias rating tool  was empl3 increase in PM2.5 and per 5 ppb increase in ozone (O3). The summary odds ratios (ORs) in the meta-analyses were estimated based on a random effects model, following the method of DerSimonian and Laird [2 statistic. The cutoff points ≤ 25%, ≤50%, ≤75%, and >75% were used to indicate no, small, moderate, and significant heterogeneities, respectively [If at lowest three studies mentioned the same air pollutant and exposure window of pregnancy available, then the impact estimates were combined in the meta-analyses. We figured summary risk estimates for congenital hypospadias per unit increase of persistent pollutant concentration. If multiple and single pollutant models were reported simultaneously, the single pollutant model analysis was chosen. In addition, if studies demonstrated effect estimate for more than one pregnancy exposure window, we extracted the results during the first trimester of gestation. To compare and contrast among different studies, units were harmonized to per 10 μg/mnd Laird . Heterogectively . Furtherectively , 54. STASupplementary Table 1Supplementary Table 2Supplementary Table 3


Defining socio-demographic factors, clinical presentations and underlying diseases associated with COVID-19 severity could be helpful in its management. This study aimed to further clarify the determinants and clinical risk factors of the disease severity in patients infected with COVID-19.A multi-centre descriptive study on all patients who have been diagnosed with COVID-19 in the province of Tehran from March 2020 up to Dec 2020 was conducted. Data on socio-demographic characteristics, clinical presentations, comorbidities, and the health outcomes of 205,654 patients were examined. Characteristics of the study population were described. To assess the association of study variables with the disease severity, the Chi-Squared test and Multiple Logistic Regression model were applied.The mean age of the study population was 52.8 years and 93,612 (45.5%) were women. About half of the patients have presented with low levels of blood oxygen saturation. The ICU admission rate was 17.8% and the overall mortality rate was 10.0%. Older age, male sex, comorbidities including hypertension, cancer, chronic respiratory diseases other than asthma, chronic liver diseases, chronic kidney diseases, chronic neurological disorders, and HIV/AIDS infection were risk markers of poor health outcome. Clinical presentations related with worse prognosis included fever, difficulty breathing, impaired consciousness, and cutaneous manifestations.These results might alert physicians to pay attention to determinants and risk factors associated with poor prognosis in patients with COVID-19. In addition, our findings aid decision makers to emphasise on vulnerable groups in the public health strategies that aim at preventing the spread of the disease and its mortalities. The Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) has now affected every corner of the world. The virus was first identified in China in the late 2019, leading to a wide variety of events including flu-like, gastrointestinal, and neurological symptoms. The disease was named as COVID-19 and was stated as a “Public Health Emergency of International Concern” by the World Health Organization (WHO) in early 2020 .Iran is one of the first and worst affected countries by SARS-CoV-2 virus, worldwide. The outbreak started from the province of Qom, where two confirmed deaths due to COVID-19 were officially reported on February 19. Shortly after, in early March, the disease reached nearly every province throughout the country [ country . Since t country .As the pandemic wears on, the health and socio-economic consequences continue to grow. Describing the clinical features and associated outcomes of patients diagnosed with coronavirus disease is crucial in improving our understanding about the disease, in optimising resource allocation to patients with the highest risk of severe outcome, and in effectively managing the pandemic. Several articles have been published worldwide, defining the characteristics and outcomes of different cohorts of patients with COVID-19 –11. HoweA descriptive epidemiological study was conducted using the registry database of Coronavirus Control Operations Headquarter in the province of Tehran. The province is located in the north-central region of Iran, and is home to the country’s capital city of Tehran. The most recent national population census held in 2016, counted a total population of 13,267,637 for the province, with 8,693,706  living in urban areas. It is the country’s most populous and important COVID-19 epicentre .In Iran, the national registry for novel coronavirus disease was established in March 2020. All suspected, probable, and confirmed case data were prospectively recorded on the national registry of COVID-19 database, by using WHO case definition guidance . In thisPatients’ data included age, sex, residing area, smoking history, opioids history, history of exposure to SARS-CoV-2, Clinical presentations , comorbidities , cancer, asthma, chronic respiratory diseases other than asthma, chronic liver diseases, chronic kidney diseases, chronic neurological diseases, chronic haematological diseases, HIV/AIDS, chronic immune deficiency diseases other than HIV/AIDS), blood oxygen saturation (SpO2%), chest CT findings, intensive care unit (ICU) admission, and the disease outcome.All variables were categorized for ease of interpretation. Patients were split into 9 age groups: 0–9, 10–19, 20–29, 30–39, 40–49, 50–59, 60–79, and 80 years and above. Sex was defined as being a woman or a man. Residing area was divided into 3 regions based on regional boundaries of three public health networks who were responsible for administration of health services in the province of Tehran. The categories were north and east, centre and south, and west. The blood oxygen saturation was classified based on the National Coronavirus Control Operations Headquarter protocol as being either higher than 93%, or 93% and lower. The disease outcome was categorized as survived, and deceased. All other variables were documented as yes or no. The history of exposure to SARS-CoV-2 was defined as having any recent contacts with a probable or confirmed case of COVID-19 from 2 days prior to the onset of the symptoms till 14 days post symptom onset, and was assessed by questioning the patient about the contact.Descriptive statistics (absolute number and percentage) were used to show the characteristics of the study population. The Chi-Squared test was performed to analyse the association of the patients’ characteristics with the blood oxygen saturation level and disease outcome. Risk factors associated with low blood oxygen saturation levels and death were explored by fitting the multivariate logistic regression model to the data. All estimates were examined by using IBM SPSS Statistics, version 26 , and the significance level was set at α < 0.05.The data was collected by trained health care professionals and based on a standardized reporting form, therefore the percentages of missing values were low and ranged between 1 to 5% across all variables used for this study. Hence, the effect of the missing data on the validity of statistical inferences was regarded as insignificant .The authors confirm that all methods were carried out in accordance with relevant guidelines and regulations, including the Declaration of Helsinki.n = 99,161) have presented with low levels of blood oxygen saturation (mean ± SD age: 58.6 ± 19.86). The ICU admission rate was 17.8%  with the mean age of 60.9 ± 20.98 for patients who required ICU admission. Overall, 20,472 individuals (10.0%) have died with the disease and the mean age of deceased patients was 67.4 ± 17.37.In this paper, data on 205,654 patients who were diagnosed with COVID-19 in the province of Tehran from March 2020 up to Dec 2020 were analysed. Of the study population, 64,468 (31.3%) were confirmed cases as defined by having positive SARS-CoV-2 PCR test result; the remaining were diagnosed according to COVID-19 associated changes in chest CT and clinically-epidemiologically criteria. Chest CT was performed for 121,973 patients, of whom 91% had shown COVID-19 associated changes. The mean ± SD age of the analytic population was 52.8 ± 21.1 years (Median 54.0 years) and 93,612 (45.5%) were women. About half of the patients  were among the children under the age of 19. The majority resided in the north and east regions of the province. Hypertension, diabetes, and cardiovascular diseases were the most common reported comorbidities. The most common symptom on admission was cough, followed by difficulty breathing, fever, and muscle ache. With regard to gastrointestinal manifestations, nausea was the most common complaint. Skin lesions and paraplegia were relatively rare conditions , male sex, residing in the north and east region of the province, having a positive history for opioids dependency, and having COVID-19 exposure history were associated with low levels of blood oxygen saturation in patients infected with SARS-CoV-2. Patients suffering from comorbidities including diabetes, hypertension, cardiovascular diseases, chronic respiratory, and chronic kidney diseases were more likely to develop low SpO2 levels. Clinical manifestations correlated with the COVID-19 related hypoxia were fever, muscle ache, difficulty breathing, diarrhea, headache, impaired consciousness, and skin lesions. The positive predictors of mortality due to COVID-19 were older age (60 and older), male sex, residing in the public health region of central and south, and having COVID-19 exposure history. Comorbidities containing cancer, asthma, chronic respiratory diseases, chronic liver and chronic kidney diseases, chronic neurological disorders, and HIV/AIDS infection were associated with higher risk of death in patients infected with SARS-CoV-2. Fever, cough, muscle ache, difficulty breathing, SpO2 ≤ 93, nausea, headache, and impaired consciousness were clinical presentations with increased odds of COVID-19 mortality. Smoking revealed a protective effect against both the low blood oxygen saturation levels and death in patients with SARS-CoV-2 infection.The mean age of patients in our study was 53 years old, which was in line with the estimated mean age of COVID-19 patients for outside China (52.5 years old) stated in a systematic review of 10 regions from around the globe. However, it was higher than the findings in China, where the mean age of patients was reported as 46 years old . CompareOur study revealed that about half of the patients with COVID-19 infection, were suffering from hypoxia at the time of admission, which was higher than the rate reported from China 36%, but lower than the rate in the United States (65%) , 22. HowIn the multivariate analysis, patients with the age of 60 and older and male sex were more likely to present with low SpO2 levels on admission, and ultimately die as the result of the disease, which confirms previous findings , 20. ResWith respect to the comorbidities associated with COVID-19 health outcome, our results revealed that patients with diabetes and cardiovascular diseases have lower odds of developing low SpO2 levels, and that the two underlying diseases were insignificantly associated with COVID-19 death. Hypertension was a positive predictor of low blood oxygen levels, but it was not correlated with death in patients with SARS-CoV-2 infection. These findings do not seem to confirm previous results which have reported these underlying diseases as important contributors of poor prognosis in patients with COVID-19. However, our results seem to be defensible since most studies have only referred to hospitalized patients , 31. AstRegarding clinical presentations, fever, difficulty breathing, and impaired consciousness were important risk factors for low blood oxygen saturation level and death. On the other hand, muscle ache, and headache were important protective factors for both conditions. Cough was insignificantly associated with SpO2 levels, but patients with cough had lower risk of death due to the disease. These results corroborate previous findings , 28. NauFinally, our study was relied on secondary analysis of existing data, hence evaluation of factors associated with the health outcomes of patients with COVID-19 were limited to available information. Given the cross-sectional nature of our study drawing conclusions about causal relationships should be done with cautious. As is the issue with most datasets on COVID-19, asymptomatic or mild cases who have not visited the healthcare facilities during the study period were not included in the analyses. Moreover, evaluation of misdiagnosis which was dependent on the sensitivity and specificity of the tests, was not possible with existing data. However, the strength of our study lies in its large multicentre study population which results in more reliable extension of inferences to the target population. Additionally, the registry from where the data was extracted, was the most complete online source of data on COVID-19 patients in the province of Tehran including all patients who have visited the public and private health care facilities in the province and were diagnosed with the disease. Given that the data was collected by trained health care professionals and based on a unified reporting online form, a high-quality data was made available to the researchers allowing real-time processing and analysis of the information.The findings of this paper indicate that older age, male sex, suffering from comorbidities included hypertension, cancer, chronic respiratory diseases other than asthma, chronic liver diseases, chronic kidney diseases, chronic neurological disorders, and HIV/AIDS infection are risk markers of poor health outcome in patients with COVID-19. The results might also alert physicians to pay attention to symptoms related with worse prognosis included fever, difficulty breathing, impaired consciousness, and cutaneous manifestations. In addition, findings about the correlates of the severe disease in patients with SARS-CoV-2 infection, aids decision makers to emphasise on vulnerable groups in the public health strategies that aim at preventing the spread of the disease and its mortalities.


This study examined the chemical, mineralogical, physical, thermal, and technological characteristics of the Dostluk (DM), Halach (HM), and Sakar (HM) clay deposits located in the Amu-Darya basin of Turkmenistan. The potential suitability of these deposits was evaluated for the local ceramic brick industry. The chemical and mineralogical features were identified by X-ray fluorescence (XRF), ion chromatography (IC), energy-dispersive X-ray spectroscopy (EDS), and X-ray diffraction (XRD) techniques. The physical properties were characterized by granulometric analysis by sieving, particle size distribution, scanning electron microscopy/optic analysis, specific surface area, Pfefferkon’s plasticity index, reabsorption, shrinkage, water absorption, mechanical (compression and bending), and freeze–thaw durability tests. The thermal methods were performed using dilatometry and thermogravimetric/differential thermal analyzer (TG/DTA). The test samples for the different clay deposits were extruded, dried, and fired at three different temperatures of 850 °C, 950 °C, and 1050 °C. While the Dostluk and Sakar clays have high plasticity, Halach clay has been found to have low plasticity. The mechanical and freeze–thaw durability tests demonstrated that the outcomes of the clays of different origins were sufficient, achieving compressive strengths of over 10 MPa and mass loss less than 3%, which are acceptable by industry standards. Semi-industrial processed hollow bricks demonstrated promising characteristics. While the Dostluk and Sakar clay-based brick specimens were visibly free of cracks, the Halach specimens showed some cracks. The physical and mechanical improvements of these clays were performed with three mixtures, which are M1 (80 mass% DM + 20 mass% brick waste), M2 (85 mass% SM + 15 mass% brick waste), and M3 (70 mass% HM + 25 mass% SM and 5 mass% brick waste) for the brick industry. The building industry has an important role in the economic development of developing countries as it is directly related to many industries, such as cement, concrete, steel and other metals, glass, tile, and bricks . One of Clays are among the most frequently used and versatile materials in industrial fields, such as the petroleum industry; they are also used in the composition of foundry molds, paint, paper coatings and fillings, pharmaceuticals, and water treatments  ,4,5,6. CThere are two main types of bricks—cement-based and clay-based bricks . Clay-baGiven these advantages, the exploration of new clay deposits in different regions of the world is important to meet the high demand for clay-based bricks from the renewable and green building industry. The Amu-Darya River is the largest river basin in Central Asia. Rising in the Pamir Mountains, it is formed by the confluence of the Vakhsh, Panj, and Kunduz Rivers. The Amu-Darya basin in Turkmenistan was studied by Brunet et al. (2017), who used geological and geophysical data to reconstruct the Late Palaeozoic and Mesozoic evolution . AccordiClays, which are natural resources, are constantly used and are subject to extinction with passing time. Waste brick (WB) is silicate industrial solid waste from either brick factories or the construction industry . The volThe main objective of this study is to examine the physicochemical features and the characterization of the ceramic bodies obtained from the Dostluk, Halach, and Sakar clays. The study evaluates the suitability of these clays using the latest diverse characterization techniques. To the best of the researchers’ knowledge, this work is the first suitability assessment of illitic Dostluk mix (DM), Halach mix (HM), and Sakar mix (SM) clays with some applications to the building industry. The study examines three different clays as raw materials to contribute to the study of the mineralogical and physicochemical features of Amu-Darya basin clays for the newly established ceramic brick industry in the Lebap region of Turkmenistan. The second objective of this work is to improve the physical and mechanical performance of clay-based bricks by using WB from this factory by making mixtures at optimum WB ratios. The output of this study provides insights into the possible methods of improving the building material features of clay-based bricks by using WB at optimum ratios.The study characterized the most suitable clays collected from three different regions of Amu-Darya basin, Turkmenistan, for the ceramic brick industry. For this purpose, deposits were selected from three distinct places, namely Dostluk, Halach, and Sakar. From a geological perspective, these three deposits are easily accessible on the edge of the Amu-Darya basin. As shown in 2. Afterward, the humidified clays were left undisturbed in sealed plastic boxes overnight. The extrusion of the clays was carried out with a 050C extruder  for specimen production in 120 × 30 × 18 mm3 dimensions. The extrusion parameters were 80 cm Hg of vacuum, 30 bar of pressure, 18 units/min of production capacity, and 33 °C to 39 °C of extrusion temperature at the exit of the extruder. The shaped samples were gradually dried in an oven at an increasing temperature between 30 °C and 110 °C until a constant mass was obtained in order to eliminate the free water content . Drying time and process varied according to the nature of the clays. If the sensitivity values obtained from Bigot’s curve are greater than 2, the wet bricks should be dried carefully. Following the drying, the specimens were finally fired at 850 °C, 950 °C, and 1050 °C within a firing cycle of 26 h. The firing cycle of DM, HM, and SM clay-based is given in After pre-treatments (drying and milling), the DM, HM, and SM clays were sprayed with about 19, 17, and 18 wt% of the total mass of the samples (6000 g), respectively, and mechanically stirred for 10 min. The amount of water required to prepare mud with these clay types was found by adjusting the penetrometer values between 1.8 and 2.5 kg/cmThe powdered clays of DM and SM were mixed with WB to make mixtures, which are M1 (80 mass% DM + 20 mass% WB) and M2 (85 mass% SM + 15 mass% WB), respectively. The mixture of HM with SM and WB clay was M3 (70 mass% HM + 25 mass% SM + 5 mass% WB). After preparing these compositions, the same procedure was followed, which was described in 2 mass. Calcium carbonate (CaCO3) equivalent was measured as follows (Equation (1)):CaCO3 = mass of CaCO3 calculated from the calibration curve (g), Mclay = mass of clay (g). The technological features of the extruded pieces were determined according to established ceramic procedures. To determine the contents of the soluble salt of DM, HM, and SM, 50 gr of clay was weighed and 500 mL of boiled water was added, stirred, and filtered. The concentration of soluble salts was detected using a Dionex™ ICS-5000+ Capillary HPIC . The calcium carbonate content of clays was determined using the Volumetric Calcimeter Method  by using calcimeter apparatus. For this, carbonates were treated with hydrochloric acid (Equations (1) and (2)) and the volume of carbon dioxide released was measured. At constant pressure and temperature, as a result of clay carbonate decomposition, the change in water level in the system is a direct measure of the resulting COd = mass of oven-dried specimens (g) at 110 °C and fM = mass of fired specimens (g) at 1000 °C. Linear drying and firing shrinkages of the specimens were determined with an indent marker stage following the test standard method ASTM C210-95. Reabsorption and water absorption were calculated based on ASTM C373-88 standard (OH)2), anorthite (CaAl2Si2O8), and gehlinite (Ca2Al2SiO7) minerals were also determined. Gehlenite and anorthite are formed by the combination of illite, silica, and CaCO3 present in the raw materials, respectively [The main stages of the firing process are evaporation, dehydration, oxidation, and vitrification, respectively . In the ectively . Mineralectively . 2O3) content [The colors of the DM, HM, and SM clay-based bricks fired at 850 °C, 950 °C, and 1050 °C were examined and found to be different from each other, as shown in  content . Althoug content . All theThe LOI test demonstrated the loss of mass at high temperatures of 850 °C, 950 °C, and 1050 °C. The LOI values of the bricks fired at these temperatures are given in Bulk density is defined as the weight (w) of a unit volume for fired brick specimens. In brief, it indicates the concentration of voids, micropores, holes, and cavities of fired bricks because these parameters determine the final density of fired bricks. As demonstrated in The apparent porosity values of the DM, HM, and SM clay-based brick specimens are given in 2 at 800 and 1000 °C, the apparent porosity decreased at 1050 °C. The apparent porosity has decreased due to the absence of carbonates in the structure of the clay used in the production of DM brick specimens. The decrease in porosity observed with increasing temperature and pressure is due to the coalescence of the molten phase and the powder compaction under pressure, respectively. After carbonate decomposition and the release of COApparent specific gravity as a significant parameter is generally expressed as the ratio of the mass of a unit volume of the impermeable part of the aggregate, which means that it does not include the permeable pores in the aggregate. The AS gravities of the brick specimens are presented in ASTM C62, the value of water absorption of less than 22% is recommended in mild weather conditions [A values demonstrate that all clays can be used in mild weather conditions, leading to sustained construction.Water absorption of fired brick specimens represents the porosity, capillaries, permeable pores, and voids in the matrix, which affect the performance of clay-based bricks . The lownditions . It has nditions ,60. Wate2 for DM, HM, and SM, respectively  are demonstrated in ectively . After fThe optical images of brick specimens  are shown in Particle morphologies and elemental analysis of the DM, HM, and SM clays are shown in 2 as a result of calcium carbonate decomposition [position . The watposition . At 1050ASTM C67, brick specimens can be considered inadmissible if the mass loss increases by 3% or if it cracks during freeze–thaw cycles [Interaction between clay-based bricks as a building material and climatic factors plays an important role in the freeze–thaw resistance of bricks . During w cycles . The iniBased on the promising features of the DM and SM clays, semi-industrial trials were carried out to test clay suitability. The results were encouraging for mass production in the newly established ceramic plant in the Lebap region of Turkmenistan. The HM bricks were also tested under plant conditions. The hollow brick samples were the first step in determining whether or not laboratory outcomes would encourage a shift from the laboratory to an industrial scale. In general, the brick specimens should not contain breaks, cracks, defects, or deformations. The brick specimens obtained under plant conditions are shown in Tons of bricks leave the factory with casualties, and these waste bricks need to be recycled .ASTM C62, all the mixtures containing WB (water absorption < 22%) are suitable for the production of ceramic bricks [It is necessary to include industrial waste in large quantities in the brick structure . Howeverc bricks . The LOIFrom the mineralogical analysis, illitic DM, HM, and SM clays were mainly composed of quartz, K-feldspar, and albite, respectively. X-ray spectrum analyses also revealed smectites in all three clays. Other minerals were also detected in DM , HM , and SM . Granulometric and size distribution analysis showed that the DM and SM clays were rich in clay-sized particles and their soil textures were clay and silty clay, respectively. Pfefferkorn analysis showed that these clay types had the highest plasticity values. HM was rich in silt-sized particles, with a prominent amount of total sand (7.34%) and quartz (33.6%). It had lower plasticity than the other clay types (DM and SM).2, MnO, and P2O3.The chemical analysis showed that the clays were rich in silica, which was over 50%. Other rich oxides were alumina, ferric oxide (resulting in the reddish color after firing), potassium oxide, and magnesia, respectively. The CaO from calcium carbonate is also observed in SM and HM. The lowest concentration oxides were TiO2O3, MgO, K2O, and Na2O) for sinterability was out of range, with an 8.90% value. The LOI values resulting from the elimination of organic matter and others for DM, HM, and SM were 7.15%, 12.40%, and 12.31% after firing at 1050 °C, respectively.The suitability plot from chemical oxides showed that the illitic DM, HM, and SM clays could be used for producing bricks, but the HM outcome from flux agents  and lower plasticity and dry-bending strength.After firing at different temperatures , all the brick specimens changed their color and sound . The bulk density, apparent density, apparent specific gravity, firing shrinkage, mass loss percentage, water absorption, freeze and thaw, bending, and compressive strengths were within the norm (standards) of building brick products. The DM and SM brick specimens had higher bending and compressive strength, while HM demonstrated the lowest strength. The strength values increased with firing temperature in all three clay types. The water absorption and freeze–thaw values of the DM and SM brick specimens were <22% and <3%, indicating that the three clays can be used for brick production. However, HM clay-based bricks displayed cracks on the edges of the bricks after 159 freeze–thaw cycles and the WA% exceeded the 22% threshold value.The industrial trials showed positive results for DM and SM, confirming the potential of DM as a raw material in the production of clay-based red bricks and SM as a raw material in the production of clay-based cream and dark beige bricks.Based on the study outcomes and observations, it can be concluded that the DM and SM clays can be effectively used for mass scale brick production, leading to energy-efficient, economical, green, and sustainable construction. HM clay could be used by mixing it with clays with high plasticity. If used alone, the brick quality will be inferior.Consequently, the WB can be used at optimum ratios, and the usage of WB at optimum ratios by mixing with clays will help to improve the brick features and minimize the negative impacts of their disposal.This study examined clay deposits from three different locations in the Amu-Darya basin of Turkmenistan to determine the suitability of these raw materials for the newly developing local ceramic industry. The DM, HM, and SM clays were characterized by mineralogical, chemical, thermal, and technological analysis to assess the brick suitability for the newly established ceramic plant in Halach, Turkmenistan. The following conclusions can be drawn from the characteristics and clay-based brick production applications of illitic DM, HM, and SM clays: