The clinical outcomes of primary immunodeficiencies (PIDs) are greatly improved by accurate diagnosis early in life. However, it is not common to consider PIDs before the manifestation of severe clinical symptoms. Including PIDs in the nation-wide newborn screening programs will potentially improve survival and provide better disease management and preventive care in PID patients. This calls for the detection of disease biomarkers in blood and the use of dried blood spot samples, which is a part of routine newborn screening programs worldwide. Here, we developed a newborn screening method based on multiplex protein profiling for parallel diagnosis of 22 innate immunodeficiencies affecting the complement system and respiratory burst function in phagocytosis. The proposed method uses a small fraction of eluted blood from dried blood spots and is applicable for population-scale performance. The diagnosis method is validated through a retrospective screening of immunodeficient patient samples. This diagnostic approach can pave the way for an earlier, more comprehensive and accurate diagnosis of complement and phagocytic disorders, which ultimately lead to a healthy and active life for the PID patients. Primary immunodeficiencies (PIDs) are a group of inherited disorders caused by defects in different components of the immune system. PIDs may be associated with severe clinical outcomes if left undiagnosed or untreated . HoweverHere, we present a newborn screening based on protein profiling that broadens the diagnosis to cover other severe forms of PIDs with innate immunity defects. The screening enables parallel diagnosis of 22 disorders due to defects in the complement system or phagocytic function prior to the onset of clinical symptoms. Complement deficiencies represent 1–29% of all PID cases,  giving aCurrently, complement functional assays, nephelometry, immunoprecipitation (IP), enzyme-linked immunosorbent assay (ELISA), complete blood count and flow-cytometric cellular characterizations are applied for diagnosis of complement and phagocytic disorders . All theThirty-seven anonymized healthy control samples in forms of dried blood spot (DBS), serum, and whole blood (including 21 newborn samples) were provided by the Karolinska University Hospital Huddinge (Sweden) for normal range identification of the targeted proteins (see Antibodies and Immunoassay Procedure sections). Moreover, 41 DBS and matched serum samples  with defects in one of the proteins C1, C2, C3, C4, C5, C6, C7, C8, C9, FB, FD, FH, FI, properdin, or HAX1 were obtained and prepared for retrospective screening . All humFilter papers (PerkinElmer) for DBS sampling were provided by the Newborn Screening Laboratory at the Karolinska University Hospital Solna (Sweden), and were prepared by applying 70 μL (corresponding to one drop) fresh and intact whole blood onto the filter paper. Papers were dried at ambient temperature (25°C) for 4 h, before desiccated storage at 4°C until use. DBS samples were punched in 3.2 mm in diameter discs (corresponding to 4.5% fraction of one blood drop), and proteins were eluted by submerging the disc in 30 μL sterile phosphate buffered saline supplemented with Tween 20  for 3 days, 1 day, or 2 h at 4°C. Subsequently, proteins from the DBS eluate or 3 μL matched whole blood were biotinylated (referred as protein labeling) by addition of ten times molar excess of Sulfo-N-Hydroxysuccinimide-polyethyleneglycol biotin  in 30 μL PBS-T (referred as labeling solution), with incubation for 2 h at 4°C. Alternatively, a rapid and simultaneous protein elution and labeling was performed by submerging the DBS disc in 30 μL of labeling solution for 2 h, 30 min, or 10 min at ambient temperature. Labeled samples were stored at −20°C until use. Immediately before incubation with the suspension bead array, 1 μL of labeled samples were further diluted 50 times in PBS buffer supplemented with polyvinylpyrrolidone, polyvinyl alcohol, and casein (Sigma-Aldrich). The diluted samples were heat-treated at 56°C for 30 min for signal enhancement  according to a previously established protocol . In brieFifty microliter of labeled, diluted, and heat-treated samples were applied to the prepared suspension bead array with bead counts of minimum 30 beads per target per measurement. After overnight incubation in a sealed plate at ambient temperature on a microplate shaker (650 rpm), the beads were washed three times in 100 μL PBS-T. The sandwich immunoassay was performed as described above, with the exception of using unlabeled DBS eluate, and addition of an incubation step with biotinylated secondary antibodies for 1 h at ambient temperature, followed by three times washes in 100 μL PBS-T . SubsequR software version 3.2.3 and R Studio environment version 0.99.902 were applied for statistical analyses and data visualization . The medInitially, we confirmed protein elution from filter paper of three healthy controls, evaluated the protein measurements, as well as testing automation and reduction in turnaround time . The proSubsequently, screening was performed on a sample cohort, including 37 healthy controls and 41 cases diagnosed with PID . The cohSamples were profiled for complement components C1-C9, complement factors FB, FD, FH, FI, and properdin, as well as the proteins related to phagocytosis and granulocyte function, including colony-stimulating factor 3 receptor (CSF3R), phagocytic oxidase subunits p22-phox, gp91-phox, p47-phox, p67-phox, and p40-phox, neutrophil elastase (ELANE) and HCLS1 associated protein X-1 (HAX1) . The defNewborn screening programs are required to meet a panel of defined criteria for the principles and practice of each diagnostic method to be considered at a public health measure. The main criteria are that the disorder needs to be a serious health issue and late-stage treatment would cause increased morbidity and mortality, while early diagnosis would lead to a significantly better outcome, a treatment or effective preventive care should be readily available for the diagnosed condition, the disorder should have a high enough incidence among the target population, the costs involved should be economically balanced to adapt for massive population screening, and the turnaround time from sampling to data should not exceed 48 h .The genetic disorders of the innate immunity are very serious, particularly when considering life-threatening infections and angioedema. Late diagnosis would cause significantly higher morbidity leading to permanent damage to vital organs and even death. A strict preventive care such as prophylactic antibody therapy, preventive treatment for systemic lupus erythematosus (SLE) and HAE, as well as postponing BCG vaccination and early hematopoietic stem cell transplantation (HSCT) in CGD patients would significantly reduce morbidity and mortality. Currently, the cumulative prevalence of deficiencies of the innate immunity, which could be markedly underestimated due to lack of accurate diagnosis, is within the range for newborn screening . The cost per assay for the proposed method is below 5 USD for the detection of the 22 disorders  and is shown to be in the range of newborn screening guidelines for massive population screening. The turnaround time from sampling to data is fewer than 10 h and fits well with the time standard.A key benefit of the developed screening system is the notable number of PID-associated proteins that can be examined in parallel using a limited amount of sample material. Moreover, the flexible design of the bead array content enables an optimal adaptation to national programs of different countries, since the prevalence and mechanism of PIDs are markedly different among ethnic groups. Nevertheless, the presented screening might miss identifying cases of rare, newly discovered types of PIDs, or disorders caused by a dysfunctional protein expression. Updating the array to include particular antibodies that bind the mutated, truncated, or distorted protein fragments might further identify these additional PID cases. The bead array technology can increase the sample throughput up to 384 individuals per instrument run, with a readout capacity of up to over 500 proteins in parallel , 20. We Taken together, combining the long-established routines for DBS neonatal sampling with the inherent potential of multiplexed technologies offers an appealing avenue that can be readily expanded to nationwide newborn screening for a broader range of PIDs. In the future perspective, a pilot program would address the aspects of population-scale screening prior to the decision of implementation in the national screening programs. In addition, the actual prevalence rates of these PIDs will be adjusted based on data from the pilot program. As an instance, the prevalence of severe combined immunodeficiency (SCID) in the US was significantly adjusted from 1 in 100,000 to 1 in 58,000 live births before and after the pilot newborn screening study . AlthougThe current study should be considered a “proof of principle” and the precursor for a pilot study with enough samples size to make a decision as to cost/benefit and feasibility. Importantly, the presented approach could pave the way for an earlier, more comprehensive and accurate diagnosis of complement and phagocytic disorders. Correct and precise diagnosis with an insight over several conditions could inform a better understanding of these diseases by finding patients with specific or combination of deficiencies, in addition to showing the differences between non-functional vs. abrogated protein expressions. With a more personalized diagnosis at hand, insights from multi-parameter assays can contribute to a more effective treatment and preventive care, which ultimately leads to a healthy and active life for PID patients.The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation, to any qualified researcher.The studies involving human participants were reviewed and approved by the Regional Ethical Review Board (EPN) in Stockholm, Sweden. Written informed consent to participate in this study was provided by the participants' legal guardian/next of kin.MD, SB, LS, HA, MN, JS, PN, and LH conceived and designed the study. MD and SB planned and performed the laboratory experiments and acquisition of data in screening. LS designed and performed the validation. MD, SB, LS, HA, MT-R, JS, PN, and LH analyzed and interpreted the data and revised the manuscript critically. LS, HA, CF, JF, CS, AR, LF, ML-T, LG-G, LA-M, YM, YY, VF, ÅL, AA, NR, AM-N, MH-G, UD, LT, TH, SN, JS, and PN contributed with patient diagnosis and/or providing of reagents, materials, analysis tools. MD wrote the paper. All authors approved the final manuscript for submission.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.


Whether the lncRNA CCAT2 expression level affects the clinical progression and outcome of cancer patients has not yet been fully elucidated. There is still an inconsistent view regarding the correlation between CCAT2 expression and clinicopathological factors, including survival data. Besides, the regulation mechanism of CCAT2 in human cancer is still unclear. Our study analyzed a large number of publication data and TCGA databases to identify the association of CCAT2 expression with clinicopathological factors and to explore the regulatory mechanisms in human cancers. We designed a comprehensive study to determine the expression of CCAT2 in human cancer by designing a meta-analysis of 20 selected studies and the TCGA database, using StataSE 12.0 to explore the relationship between CCAT2 expression and both the prognosis and clinicopathological features of 33 cancer types and 13285 tumor patients. Moreover, we performed GO and KEGG pathway enrichment analyses on potential target genes of CCAT2 collected from GEPIA and LncRNA2Target V2.0. The level of CCAT2 expression in tumor tissues is higher than that in paired normal tissues and is significantly associated with a poor prognosis in cancer patients. Besides, overexpression of CCAT2 was significantly associated with tumor size, clinical stage, and TNM classification. Meanwhile, CCAT2 expression is the highest in stage II of human cancer, followed by stage III. Finally, 111 validated target gene symbols were identified, and GO and KEGG demonstrated that the CCAT2 validation target was significantly enriched in several pathways, including microRNAs in the cancer pathway. In summary, CCAT2 can be a potential biomarker associated with the progression and prognosis of human cancer. The rapid advancement of exploring ncRNA is the result of RNA-Seq technology, which provides a deeper understanding of the human transcriptome. Further research on these RNAs will lead to new perspectives on cancer cell regulation mechanisms and innovative therapeutic targets . AccordiCCAT2 is located in the 8q24 gene desert. The locus was first named in colon tumor tissue in the 2000s . OverexpThe above evidence indicates that CCAT2 is involved in tumor progression. Moreover, some previous meta-analyses have reported that increased CCAT2 expression is significantly associated with lymph node metastasis, distant metastasis, and higher clinical stage , 14. HowWe searched the English and Chinese medical literature in PubMed, Web of Science, Wiley Online Library, Weipu, Wangfang Data, and CNKI to identify all publications related to CCAT2 in cancer patients. The search strategy combining the terms “CCAT2” or “colon cancer associated transcript 2” is based on the purpose of the study. We also reviewed comments and references related to CCAT2 by other methods, including the extraction of previous articles cited in the meta-analysis of CCAT2 –15. The The meta-analysis study was evaluated by two independent investigators (RG Guan and D Liu) using the same multistep approach. First, check the headlines and abstracts to exclude unqualified studies that are not relevant, duplicate papers, reviews, or case reports. What is more, the full text of the remaining studies was further examined separately by the same investigator (M Yu). Finally, the third commentator (BW Huang) resolved any disputes.We consider studies that meet the following inclusion criteria to be eligible: (i) collecting clinical samples from tumor tissue, (ii) studying the association between CCAT2 and survival data and performing CCAT2 levels by qRT-PCR quantification, and (iii) providing sufficient data to determine the HR value and its 95% CI. The exclusion criteria were as follows: (i) repeated studies, (ii) study data not sufficient to calculate HR values, (iii) studies of animals or cell lines, and (iv) reviews, comments, letters, case reports, and conference articles. If the survival analysis is not sufficient to calculate the HR value, we try to contact the author to obtain the raw survival data.https://github.com/markummitchell/engauge-digitizer/releases).The following relevant information from all eligible studies was extracted: first author name, publication year, country, cancer type, sample size, assay method, the criterion for dividing CCAT2 into high and low groups, follow-up time, prognostic data, age classification, gender ratio, tumor size, clinical stage, TNM classification, and histological differentiation. If the paper does not provide complete survival data, we follow the methods of He et al. . The HR The NOS was used to assess the methodological quality of two investigators (RG Guan and D Liu) independently evaluating eligible projects. They rated each study according to the following system: (i) selection, 0-4; (ii) comparability, 0-2; and (iii) exposure, 0-3. The highest score is 9 points, and score ≥ 6 indicates that the research quality is good.Q test and the I2 index were used to assess potential heterogeneity in selected studies, with P′ < 0.05 or I2 > 50% considered statistically significant. If the selected parameter has significant heterogeneity (P′ < 0.05), the random effects model is used to calculate the HR value; otherwise, a fixed effects model will be employed. Finally, Begg's test was used to estimate publication bias .We used StataSE 12.0 software to analyze the information extracted from eligible studies. HR and 95% CI assessed survival data. The OR and 95% CI were calculated to analyze the relationship between human cancer and clinicopathological parameters, including age, gender, tumor size, clinical stage, TNM classification, and histological differentiation. What is more, subgroup analyses were based on the source of tumor type and overall survival data. The Cochrane http://starbase.sysu.edu.cn/). Experimental data were divided into high and low groups based on the median level of CCAT2 expression. The Cox proportional hazard model of SPSS 22.0 was used to assess the effect of CCAT2 overexpression on survival. The box diagram and bar graph of CCAT2 expression in tumor samples and adjacent normal tissues were drawn by R 3.6.0 (https://www.r-project.org/) and established on the data extraction of the TCGA database .CCAT2 expression levels and overall survival data in the TCGA database were extracted from starBase (http://gepia.cancer-pku.cn/) and LncRNA2Target V2.0 (http://123.59.132.21/lncrna2target/index.jsp) based on all published lncRNA papers to identify potential CCAT2 target genes in human cancer. GO enrichment analysis and KEGG pathway analysis were performed using DAVID Bioinformatics Resources 6.8 (https://david.ncifcrf.gov/). We used R 3.6.0 to visualize the results of GO and KEGG and used Cytoscape 3.7.1 (https://cytoscape.org/) software to display a network of CCAT2 and its related genes.We used GEPIA (n = 2192) were included in our analysis, and the main characteristics are shown in We researched to analyze the connection between CCAT2 expression and prognosis in cancer patients in all published literature . A totalP = 0.036, I2 = 70.4%, and P′ < 0.001)        (As shown in < 0.001) , clinica= 0.238) , T stage= 0.347) , N stage= 0.009) , and M s= 0.078) . However= 0.741) , gender = 0.996) , and his< 0.001) . The abon = 2192) and the TCGA database (n = 11093), suggesting that CCAT2 overexpression is significantly associated with poor prognosis for certain cancer types. Considering the significant heterogeneity in the study, we performed two subgroup analyses based on survival data and the source of the cancer type. In a subgroup analysis of OS, we found that high expression of CCAT2 was significantly associated with poor OS in all databases , including publications  and TCGA  (P < 0.001), osteosarcoma , cholangiocarcinoma , gastric cancer , breast cancer , and others )  . As pitu 0.001)) . No signP = 0.496) and TCGA (P = 0.455) (Figures Begg's funnel plot was used to assess publication bias in our study. No publication bias was observed in studies evaluating the association of CCAT2 with clinicopathological features and OS in the study group ( Figures . Similar Figures . Among tBased on the results obtained from TCGA, we plotted a box diagram of the CCAT2 expression profile for tumor samples and adjacent normal tissues . We founTo explore the underlying mechanism of action of CCAT2, we identified a total of 111 target genes using GEPIA and LncRNA2Target V2.0. GO and KEGG analysis was performed. CCAT2 and target gene symbols were analyzed by GO enrichment analysis, including BP, CC, and MF, and the results are shown in Numerous studies have shown that overexpression of CCAT2 is significantly associated with clinical outcomes and other clinicopathological parameters in cancer patients –18. The A meta-analysis of 2192 patients from 20 eligible studies and 11093 patients from TCGA currently explores the association between CCAT2 overexpression and prognosis, as well as the clinicopathological parameters of cancer patients. Therefore, our research is by far the most comprehensive analysis. We assessed the quality of all selected studies through NOS and used Begg's method to examine publication bias. Our results show that high expression of CCAT2 is associated with poor OS. For clinicopathological features of cancer patients, our study suggests that high CCAT2 is significantly associated with cancer growth and metastasis, including tumor size, clinical stage, and TNM classification, although age, gender, and histological differentiation are not significant factors. The results suggest that CCAT2 may be a potential tumor biomarker and is associated with tumor invasiveness, which is why CCAT2 is mainly expressed in stage II, followed by stage III.Furthermore, a subgroup analysis of CCAT2 expression and overall survival was not statistically significant in TCGA, and CCAT2 is likely overexpressed in certain types of tumors. Besides, subgroup analysis was also performed on specific cancers, including hepatocellular carcinoma, osteosarcoma, cholangiocarcinoma, gastric carcinoma, and breast cancer. Increased CCAT2 expression was associated with worse HR observed in hepatocellular carcinoma, cholangiocarcinoma, gastric cancer, and breast cancer, whereas no significant association between CCAT2 expression and HR was detected in osteosarcoma. However, KIRC, PRAD, READ, SKCM, and STAD in the TCGA data set are associated with a good prognosis. We reviewed related studies and found that overexpression of CCAT2 levels is associated with worse outcomes in renal cell carcinoma , prostatStudies have shown that CCAT2 expression levels are upregulated in cancerous tissues compared to paired adjacent tissues; the same results were found in in vitro cell line samples . ResearcThe analysis of GO and KEGG pathways suggests that CCAT2 may play a key role in human tumors through different pathways, including miRNAs in the cancer pathway, etc. miRNAs are defined as small noncoding sets of 19 to 24 nucleotides associated with mRNA expression and regulate the expression of downstream gene targets, including oncogenes, tumor suppressor genes, and transcription factors . StudiesCompared to previous meta-analyses, our research has several advantages –15. FirsAlthough our study attempts to fully elucidate the association between CCAT2 and cancer progression and prognosis, our research has some limitations. For the meta-analysis, different definitions of high CCAT2 expression levels in selected studies are factors that contribute to publication bias. Besides, the current eligible countries in the meta-analysis are only China, the USA, and Japan, and more trials in other countries should confirm our research. At the same time, since there is no direct data for multivariate analysis in some existing studies, we have to extract relevant data through Kaplan-Meier curves, which may lead to deviations in HR values. More importantly, all available studies are retrospective studies that tend to be published when positive results are confirmed. Thus, the impact of CCAT2 on the prognosis and clinicopathological parameters of malignant tumors may be overestimated. Therefore, further research is needed to study the clinical significance and diagnostic value of CCAT2 in human cancer. Furthermore, although CCAT2 can act through a variety of mechanisms, based on the correlation of gene expression levels between CCAT2 and miRNA, only one possible mechanism of the role of CCAT2 in gene regulation has been investigated. In order to understand more features of CCAT2, further research is needed to explore other possible mechanisms.In light of our findings, we believe that the expression of CCAT2 may serve as potential candidates for prognostic factors as well as therapeutic targets in malignant tumors. Of note, the prognostic roles of CCAT2 varied greatly across cancers, which implied a noteworthy amount of heterogeneity between different types of tumors. In addition, the expression of CCAT2 was closely associated with tumor size, clinical stage, and TNM classification and mainly expressed in stage II, which indicated that CCAT2 is a significant biomarker to monitor tumor progression. Current findings enhance our understanding of the CCAT2 in cancer monitor and identify strategies for the early invention in clinical management. Moreover, further illumination of the underlying mechanism and the interaction between CCAT2 and tumors may provide important implications for the success of early monitoring and prognosis prediction in cancers.Our study demonstrated that a higher CCAT2 expression was significantly associated with an aggressive disease course in patients with cancer, predicting a larger tumor size, more advanced clinical stage, more inferior TNM classification, and shorter OS. We also demonstrated that CCAT2 plays an essential role in the biological processes of tumor progression via a variety of pathways.


BMAL1, CRY2, NR1D1, NR1D2, PER2, RORA, RORC, and TIM genes in persistent AF group than in other groups. There was a significant correlation between higher AHRE burden and larger LA size and between higher AHRE burden and decreased expression of circadian clock genes in patients with AF. LA volume and the expression of CRY1, NR1D1, and RORA are significantly associated with AHRE burden. However, the underlying mechanism needs to be elucidated in further studies.A prominent circadian variation is present in atrial fibrillation (AF) attacks that may be related to the expression of circadian clock genes. Little is known about the expression of circadian clock genes in AF. We prospectively enrolled 73 patients who had received pacemaker implantation, in order to define the burden of atrial high-rate episodes (AHREs) accurately. AF was diagnosed clinically in 43 (59%) patients . The expression levels of circadian clock genes of peripheral blood leukocytes were checked. There were more males and patients with a larger left atrial (LA) size and lower expression levels of  BMAL1, CK1ε, CLOCK, CRY1, CRY2, NRID1, NR1D2, PER1, PER2, PER3, RORA, RORB, RORC, and TIM. A major feature of circadian rhythm is that the mRNA and protein of the circadian clock genes and the mRNA and protein of the genes regulated by circadian rhythm exhibit distinct daily day–night fluctuation cycles , NR1D1 , RORA , and LA volume  were significantly associated with the AHRE burden    . The stu < 0.05) .CRY1, NR1D1 and RORA were significantly associated with AHRE burden. Third, there was a significant correlation between LA size and decreased expression of circadian clock genes.There are several important findings in this study. First, patients with persistent AF had a decreased expression of circadian clock genes compared with the paroxysmal AF and no AF groups. Second, LA volume and the expression of Many studies have reported the relationship between AF and sleep disorders, including insomnia and obstructive sleep apnea syndrome . Sleep dPER3 tandem repeat exhibited an elevated heart rate, and selective deletion of PPAR-γ, a putative activator of BMAL1, in the vasculature resulted in diminished heart rate diurnal variations [BMAL1 and PER3 was strongly associated with AHRE burden, which conforms to the findings of previous studies. Our study revealed that the expression of BMAL1, CRY2, NR1D1, NR1D2, PER2, RORA, RORC, and TIM was also associated with AHRE burden, which implies that other circadian clock genes may also play an important role in the variability of heart rate change, and then in the development of AF. Further studies are needed to investigate the role of circadian clock genes changes in the occurrence of atrial tachyarrhythmias, including AF.At present, diurnal changes in the human heartbeat are considered to be related to certain circadian clock gene fragments . In prevriations ,21. The riations ,23. Our BMAL1 or NRID1 in macrophages enhanced cytokine production and diminished their rhythmic expression [Macrophage-produced cytokines, including interleukin (IL)-l, IL-6, IL-12, and tumor necrosis factor-α (TNFα), fluctuate in a circadian pattern and generally peak at the end of the rest period. A previous study showed that deleting pression . Sleep dpression . These mpression . AF coulpression . Anotherpression . It is pAnother interesting finding is that some circadian clock genes expression were slightly increased in patients with paroxysmal AF, and then were significantly decreased in those patients with persistent AF. One plausible explanation is that the increasing circadian clock genes expression may have been a response to AF triggering factors  or tachyarrhythmia itself, and the response flattened or even depressed under a longer duration of stress and/or tachyarrhythmia. Another possible explanation would be that the different circadian clock genes expression patterns that represent the mechanism of circadian clock genes alterations, resulting in or caused by different AF types , may be different. Further studies on the relationship between circadian clock genes expression and AF type are needed to clarify the underlying mechanism.Our study also found there was a strong, positive correlation between higher AHRE burden and larger LA size, and that there was also a strong, positive correlation between higher AHRE burden and decreased circadian clock genes expression in patients with AF. Although AHREs are not equivalent to AF and also includes other atrial tachyarrhythmias , previous studies have shown that a higher AHRE burden was significantly associated with the development of AF ,30. The CRY1, NR1D1, and RORA, in addition to LA size, can predict AHRE burden during follow-up. PER and CRY protein accumulate in the cytoplasm and form a heterodimer and translocate to the nucleus to inhibit the transcriptional activity of BMAL1-CLOCK (negative feedback loop). NR1D1 and RORA form the secondary feedback loop and act as repressor and activators of BMAL1 transcription [Most important of all, the expression of cription . Our stuAccording to previous studies, LA size is not only associated with AF duration, but is also an independent risk factor for AF and a predictor of recurrence of AF after therapy . With thThere were several limitations in this study. First, we could measure the expression of genes during the daytime only. However, the significant down-expression of several genes at the same time point in persistent AF patients, compared to those with paroxysmal AF or no AF, also implies that there is a link between the disruption of circadian rhythm and persistent AF. Second, other factors, such as diet and light exposure, are already known to influence the expression of circadian clock genes. In the present study, we asked participants to maintain a regular lifestyle before blood sampling, to decrease the influence of these factors on circadian clock genes expression. However, we did not have the survey of well-being, general health . Third, this is a study with a smaller sample size event though it already achieved statistical significance. This study may be considered as a pilot study and further larger cohort studies should be performed to confirm the finding. Fourth, the purpose of this study was not to diagnose or predict AF. Therefore, we did not have the sensitivity, specificity, and predictive value of each respective genes. Although our results may not be useful in enhancing the diagnostic ability for AF or AF burden, we would more like to conclude the expression of circadian clock genes are associated with AF type, AHRE burden, and LA size. In addition, the LA size and the expression of some of the circadian clock genes could be used to predict higher AHRE burden in patients with AF. In those AF patients with potential higher AHRE burden, more aggressive ECG monitor and even implantable cardiac device may be indicated to document the AF or AHRE burden and even used for risk stratification. Fifth, we revealed the relationship between the expression of circadian clock genes and different AF types only, and not the causal effects. Further studies investigating the underlying mechanism between circadian clock genes expression and AF are warranted. Finally, we did not analyze the relationship between medication for AF treatment and the expression of circadian clock genes. There are several known regulators of circadian function, such as glucocorticoids, 2-methoxyestradiol, forskolin, PKC and p38 MAPK inhibitors. However, in the current literature to date, no known drugs used to treat AF will affect the expression of circadian clock genes.CRY1, NR1D1, and RORA were significantly associated with AHRE burden. However, the underlying mechanism needs to be elucidated in further studies.Our study showed that the mRNA expression of circadian clock genes was decreased in patients with persistent AF, compared to those with paroxysmal AF or no AF. The altered expression of circadian clock genes was correlated significantly with a higher AHRE burden and a larger LA size. LA volume and the expression of


Paget disease is a complex disorder that can be identified in the breast (mammary Paget disease) or in other locations (extramammary Paget's disease) such as ano-genital skin (Paget disease of the vulva -PVD). This condition is associated with low mortality, but a late diagnosis and recurrence can negatively impact the prognosis. Therefore, the main objective of this study is to evaluate if the human epididymis protein 4 (HE4) and cancer antigen125 (CA125) can promote recognition of PVD in early stages and during the relapses.we have conducted a prospective, observational and laboratory-based study, that included 50 patients, whose 25 healthy women represented the control group and 25 PVD patients, which have been operated in our Oncology Institute, from May 2017 to September 2019. Both in the control group and in PVD patients, the CA-125 and HE4 were evaluated before surgery and after 6 months. Finally, a comparison of markers serum level, both between before/after surgery and with control group, and a ROC (Receiver Operating Characteristic) curve were performed.P = .3) and HE4 (P = .19). On the other hand, comparing HE4 in PVD patients with the control group, a statistically significant difference was found . Contrary, comparing CA-125 in PVD patients with the control group , no statistically significant difference was evidenced. Moreover, ROC (Receiver Operating Characteristic) curve showed low sensitivity and specificity for CA125 with area under curve (AUC) = 0.5608. Instead, the ROC curve of HE4 revealed a sensitivity and specificity of 76% and 88% respectively (AUC = 0.7408) using a cut-off at 90 pmol/L.Dosing the markers in PVD patients, 3/25 (12%) showed a higher value of CA125 and 11/25 (44%) an increased HE4. In addition, after surgical treatment there were no statistically significant difference between levels of CA-125 (Despite the limited cases, our data showed that CA125 is not a sensitive marker for PVD. On the other hand, in 44% of PVD we’ve seen an increase in HE4. So, this could be a starting point for further research that could confirm the possibility to use this marker in order to support PVD early identification. HE4 is only slightly expressed at the epithelium level of respiratory and reproductive organs, while it is over-expressed in ovarian cancers. Therefore, it has been suggested as a complementary or alternative serum marker to carbohydrate antigen 125 (Carbohydrate Antigen 125 -CA125) for ovarian cancer (OC) risk assessment in presence of ovarian neoformation. Several studies demonstrate that HE4 has a sensitivity similar to CA125 and a greater specificity in patients with OC, while CA125 is reported in multiple physiological and pathological conditions such as pregnancy, menstruation, endometriosis. Whereas, the combination of CA-125 and HE4 in ovarian pathology, showed a sensitivity of 76.4% and a specificity of 95%. Despite, the undisputed validity of these markers in OC patients, the potential use in other types of cancers is still under investigation. In particular, our goal is to evaluate the variation of CA125 and HE4 in patients with diagnosed of Paget disease of the vulva (PVD). Paget disease represents a rare cancer , most commonly found in postmenopausal women (Caucasian ethnicity), that can be located in the breast (mammary Paget disease) or anogenital area.Human Epididymis Protein 4 (HE4) is a secreted glycoprotein, encoded by the omolologist gene and with the coexistence of invasion areas (adenocarcinoma) characterized by the presence of Paget cells infiltrating the underlying dermis. At the vulvoscopic examination, PVD is identified for the presence of a red and white eczematoid plaque, with a papillomatous and sometimes ulcerated surface and is often associated to irritation, itching, burning and vulva pain but without any pathognomonic symptoms. Consequently, PVD is often diagnosticated only in presence of very extensive disease with consequently indication to a demolition surgery and with a high percentage of local recurrence (30%–35%). Therefore, the aim of this study is to assess the effectiveness of CA-125, HE4 markers, to value the presence of PVD in women with a vulvar lesion. On the other hand, a secondary endpoint is to compare marker dosage in PVD patients, 6 months after surgery and in relation with group control.(Extramammary Paget's disease). Furthermore, 54% of PVD could be associated with lesions in other places 2 for HE4, the predetermined thresholds in menopausal patients’ status, are less than or equal to 140 pmol/L.,14 Serum markers results and the histopathological analysis were received after surgery, by a gynecologic pathologist and the pathological diagnosis of PVD has been identified by 2 pathologists of our Institute and has been defined as PVD “Invasive” the extension for at least 1 mm beyond basal membrane while the “positive margin” has been defined in case of the presence of cells within 1 mm of the surgical margin. During clinical follow-up  the tumor markers dosage was repeated and the comparison of serum levels before/after-surgery and with control group was performed.This is a prospective, observational, laboratory-based study, concerning the dosage of biomarkers on peripheral blood of 50 consecutive patients submitted, of which 25 patients  with histological diagnosis of PVD and surgically treated by Gynecologic Oncology Unit in National Cancer Research Centre, Istituto Tumori “Giovanni Paolo II” of Bari between 2017 and 2019. Menopause status was defined as the absence of menstruation for more than 6 months, the presence of clinical signs of menopause, or with a history of hysterectomy. On the other hand, 25 healthy voluntary subjects, with similar characteristics , who usually perform the periodic follow-up  for oncologic prevention, were enrolled. Consequently, in the healthy volunteers, a blood sampling for the dosage of CA-125 and HE4 was performed, after obtaining informed consent to participation in the study and to use personal data reported on specific database. Similarly, during a pre-inclusion visit of 25 patients with PVD, all demographic information, anamnestic, histological and diagnostic examination  have been collected and reported on specific database. All participants have been properly informed about research protocol, approved by the Institutional Review Board and subsequently they have also signed an informed consent both for participation of the study and for the use of personal data. In addition, on the day of surgery, during the preoperative evaluation, a blood sample was collected to perform CA125 and HE4 assays. In order to minimize variable effects due to sample collection, processing, and storage temperature, all blood samples were managed in the same manner without any protocol amendment during the entire collection period. The blood specimens were collected in serum separation tubes , allowed to clot for thirty minutes, and then centrifuged at 3500 rpm for 15 minutes for the subsequent routinely use. The HE4 and CA125 levels were detected before and after surgery with an immune-enzymatic assay (Roche Diagnostics S.p.A.) according to manufacturer's guidelines. Concerning CA-125, the reference values are less than 35 U/mL,3P value = 0.3698) and HE4  after surgical treatment and also comparing CA-125 of PVD patients with the control group . Moreover, in the control group, 6/25 patients (24%) had CA-125 levels above the cut-off, 1/25 (4%) had HE4 levels higher than cut-off. No statistically significant differences were observed comparing before and after CA 125 and HE4 values (data not shown). ROC Curve showed low sensitivity and specificity for CA125 (AUC = 0.561) while the ROC curve of HE4 revealed a sensitivity and specificity of 76% and 88% respectively (AUC = 0.7408) using a cut-off at 90 pmol/L  presented a higher value of CA125 and 11/25 (44%) an increased HE4. In addition, was not recognized a statistically significant difference of CA-125  curve and relative AUC (Area under curve) were calculated both for CA125 and HE4. Statistical analyses were performed using Graph pad Prism 5.0 software.In order to compare CA125 and HE4, before and after surgery and with control group, the Kruskal-Wallis while t-test was used. The level of statistical significance has been set to 4 On the other hand, the applying of specific reactive (acetic acid and Lugol's iodine), commonly adopted in cervical cancer screening, is not indicated for vulvar lesions evaluation. Consequently, the use of non-invasive procedure as markers serological dosage (HE4 and CA125) to support the diagnosis could be extremely helpful in order to guide the early PVD diagnostic-therapeutic process and the identification of recurrences. This is especially proper, regarding a rare disease as PVD, whose clinical knowledge are limited and the clinical interpretation may be equivocal.–18 Indeed, the differential diagnosis includes skin candidiasis, seborroic dermatitis, psoriasis, Bowen disease and melanoma. Therefore, since an exceptional number of PVD cases have come to the observation of our clinic and some of these had an increase in HE4, that is expressed also in epithelial tissues, we tried to establish if this recently proposed biomarker could be associated with the presence of PVD and consequently suitable in PVD diagnosis and/or management. Our data shows that, using the assessed HE4 cut-off (140 pmol/L), 44% of patients with PVD have a higher HE4 value and compared with HE4 dosing in the control group a statistical difference was found. Consequently, this marker could direct the clinician to perform a vulvar biopsy in case of suspected lesion and during the follow-up. In contrast, CA125 evaluation, seems to be not indicated in the presence of PVD. This assessment is further confirmed by the absence of a significant difference of CA125 both after surgery and compared to the control group. Moreover, the ROC analyses of HE4 highlighted some suggestion to be discussed. In particular, by lowering the cut-off threshold from 140 pmol/L to 90 pmol/L, the sensitivity improved greatly from 44% to 76% with an acceptable specificity of 88%.PVD could be diagnosed after a vulvoscopic examination, which is usually performed through a colposcopic or a dermatoscopic inspection. Indeed, a significantly higher values of CEA in affected patients compared to control groups was found. Nevertheless, even in the latter case, it is far from establishing the real utility of this biomarker and the potential introduction in clinical practice.Nerveless, concerning PVD and oncological markers, particularly HE4, no data are presented in literature, so it is difficult to compare our result. On the other hand, a recent study reports the assessment of tumor markers in vulvar cancer, showing that the best diagnostic performance was achieved for Carcinoembryonic Antigen (CEA).5PVD can remain undiagnosed for several years, so frequently it is recognized as an extensive vulvar lesion which needs the use of demolition surgery and subsequent plastic-reconstruction.Therefore, the search for serological markers to assist the early detection of PVD, would allow the identification of limited and non-invasive forms and the use of alternative approaches such as imiquimod and photodynamic treatment (currently off label).Actually, none of the markers analyzed are helpful in the specific identification of PVD, but the increase HE4 value, in vulvar lesion, could support clinician decision to perform a biopsy and early detection of PVD that consequently could improve the mortality and morbidity.It is also necessary to consider limitations of this study, because of restricted number of cases and for the data absence in the available publications concerning the association between PVD and serological marker. Therefore, this experience could be a valid tool to be used in routine clinical practice and possibly, a cornerstone for further discussion on the topic also considering the rarity of this pathology. It also may provide useful recommendations for national and international gynecological society.This research project has been supported in part by the Apulian Regional Project “Medicina di precisione”.Data curation: Vera Loizzi.Formal analysis: Carla Minoia, Gennaro Cormio.Funding acquisition: Carla Minoia, Angelo Paradiso.Investigation: Erica Silvestris.Methodology: Porzia Casamassima, Vito Michele Garrisi.Project administration: Antonio Tufaro.Resources: Skrypets Tetania.Supervision: Giulio Gargano, Carmine Carriero, Angelo Paradiso, Gennaro Cormio.Validation: Gennaro Cormio.Visualization: Vera Loizzi, Gennaro Cormio.Writing – original draft: Miriam Dellino, Vito Michele Garrisi.Writing – review & editing: Raffele Tinelli.


Exposure to environmental noise has been associated with an increased risk of cardiovascular diseases and diabetes, but evidence for occupational noise is limited and conflicting, especially related to pregnancy outcomes. This study aimed to evaluate the association of occupational noise exposure with hypertensive disorders of pregnancy (HDP) and gestational diabetes.Our population-based cohort study utilized data on 1 109 516 singletons born to working mothers in Sweden between 1994–2014 from the Medical Birth Register and the Longitudinal Integration Database for Health Insurance and Labor Market Studies. Noise exposure came from a job exposure matrix (JEM) in five categories <70, 70–74, 75–80, 80–85, >85 dB(A). Relative risks (RR), adjusted for confounders and other job exposures, were calculated by modified Poisson regressions for the full sample and a subsample of first-time mothers reporting full-time work.Exposure to 80–85 dB(A) of noise was associated with an increased risk of all HDP  and preeclampsia alone  in the full sample. Results were similar for first-pregnancy, full-time workers. Exposure to >85 dB(A) of noise was also associated with an increased risk of gestational diabetes  in the analysis restricted to first-time mothers working full-time.In this study, exposure to noise was associated with an increased risk for HDP and gestational diabetes, particularly in first-time mothers who work full-time. Further research is needed to confirm findings and identify the role of hearing protection on this association so prevention policies can be implemented. Hypertensive disorders of pregnancy (HDP) – which include gestational hypertension, preeclampsia, and eclampsia – and gestational diabetes mellitus, are some of the most common conditions during pregnancy, affecting between 1–15% of pregnancies throughout the world . HDP canApproximately one quarter of all hypertension cases during pregnancy will progress to preeclampsia; however, predicting who progresses has proven difficult . The cauSeveral studies have linked environmental noise exposure to cardiovascular disease –11 and dThe aim of this study is to explore whether noise exposure during pregnancy is associated with HDP, looking specifically at women who develop preeclampsia alone, as well as gestational diabetes, using a large, register-based, prospective dataset, adjusting for other occupational exposures and potential cofounders.A prospective cohort was formed based on information on each pregnancy from the Swedish Medical Birth Register for all births between 1994–2014. In this register, covering approximately 99% of all pregnancies , data arThis register was merged with the Longitudinal Integration Database for Health Insurance and Labor Market Studies (Swedish acronym LISA) utilizing the unique personal identification number assigned to all individuals living in Sweden in order to obtain information on socioeconomic characteristics, such as mother’s highest education at the time of delivery and marital status. LISA has complete coverage of all individuals aged 16 and older residing in Sweden.To examine noise exposure, we used a job exposure matrix (JEM) linked to the cohort data using occupational codes. Occupational information provided at the prenatal care unit was recorded as free text, which was then coded into the ISCO-88-based standard for Swedish occupational classification 96 (SSYK96) coding system by an occupational hygienist. The coding details are presented elsewhere . SSYK96 The noise exposure job matrix includes information on annual average 8-hour occupational noise level in decibel [dB(A)] in five-year intervals from 1970–2014 for 321 occupations. Noise levels were derived from measurement reports from occupational health services and clinics as well as large companies throughout Sweden and were specified in five categories: <70, 70–74, 75–80, 80–85, >85 dB(A) . A previSome occupations did not have enough detail to be given a 4-digit SSYK96 code and were given a 2- or 3- digit code; thus, they could not be matched directly with the JEM job groups. For these cases, imputation was done in one of two ways. First, if all occupations within this 2- or 3-digit job groups had the same noise category, that category was imputed for the missing observations. For instance, all occupations within the ‘manager of small companies and entities’ job group had the lowest noise level; therefore, women who were coded as belonging to this overall job group were also given the same level. Second, if the noise levels varied within the 2- or 3-digit job group, an occupational hygienist with expertise on noise exposure and blinded to the diagnoses of individuals within each group was consulted, and he then assigned the most likely noise level for those observations.For each mother–child pair included in our study, we extracted diagnoses coded at the end of each pregnancy reported in the Medical Birth Register. These were coded using the International Classification of Diseases, ninth and tenth revisions (ICD-9 and ICD-10). For HDP, ICD-9 codes 642, 642D, 642E, 642F, 642G, and 642H and ICD-10 codes O11, O13, O14.0, O14.1, O14.2, O14.9, O15.0, O15.1, O15.2, and O15.9 were used. For preeclampsia, ICD-9 codes 642E, 642F, and 642H and ICD-10 codes O11.9, O14.0, O14.1, O14.2, O14.9, O15.0, O15.1, O15.2, and O15.9 were used. Finally, for gestational diabetes, ICD-9 codes 648A and 648W and ICD-10 codes O24.4 and O24.9 were used.2), normal weight (18.5–25 kg/m2), overweight (25–30 kg/m2), obese (≥30 kg/m2)], country of birth , parity , and employment status . Occupational exposures considered were whole body vibrations, any particle exposure, physically strenuous work, job strain, and exposure to low temperatures, which were obtained from various JEM and matched to each woman based on occupational code and year, if applicable.Several potential confounders were considered. From the Medical Birth Register we obtained maternal age at delivery , smoking status at week 10 , educational level , marital status , family situation , body mass index (BMI) calculated using height and weight [underweight (<18.5 kg/m2. The occupations that did not have enough detail (coded on a 2- or 3-digit SSYK96) were those in which all sub-groups (coded on a 4-digit SSYK96) were not exposed to vibrations; therefore, all were assigned a value of 0. We also considered occupational exposure to particles by including a JEM based on the Finnish Information System on Occupational Exposure JEM . Covariate adjustment followed a two-step procedure. First, we selected separate sets of confounders for each analytical model based on known risk factors for HDP and gestational diabetes , 7, placTo explore multicollinearity between the exposure and confounders, we included them into a linear regression model and calculated the variance inflation index (VIF). VIF values were well <5 for all categories of all covariates, indicating that, statistically, there were no major issues with multicollinearity.Because different births from the same mother could not be assumed to be independent, relative risks (RR) were estimated using a modified Poisson regression for correlated binary data . Crude aThis study was conducted with approval from the regional ethical review board in Stockholm, Sweden.A total of 1 109 516 mother-child pairs were included in the study. The characteristics of the cohort are described in Results from the regression analyses for the full sample, which includes both full- and part-time workers, as well as first and subsequent pregnancies, are found in We conducted a separate analysis restricting to women who were in their first pregnancy and working full-time, the part of the full sample who were more likely to be present at work and thus more likely to be exposed to the levels within the assigned noise category. This resulted in a subsample of 349 300 pregnancies; results are presented in In this nationwide cohort study, exposure to noise at the workplace was associated with an increase in HDP and preeclampsia, notably for first-time pregnant women who worked full-time, the subsample of the full cohort we believe are more likely to be truly exposed. These associations were present beginning at 80 dB(A); however, were not statistically significant at levels >85 dB(A). Occupational noise exposure was further associated with an increase in gestational diabetes among first-time pregnant women working full-time who were exposed to noise levels >80 dB(A).Previous studies have found conflicting results regarding noise exposure and pregnancy outcomes. One environmental study reported an increased risk for severe and early onset preeclampsia but not for mild and late onset . Three oOur results for the 80–85 dB(A) category were largely driven by the childcare workers, which make up the majority of this group. Because data for this study come from a birth register with routine data collection, we do not believe that women who work in childcare and have a pregnancy outcome are disproportionately represented in the dataset. Restricting the sample and removing child care workers gives the similar results as in the full sample, except for the sub-group of restaurant and kitchen workers, whose inclusion did not show an increased risk (data not shown). Further studies should be conducted to elucidate these findings.The subsample analysis of women who were in their first pregnancy and working full-time showed a similar effect for both HDP and preeclampsia. Preeclampsia is generally considered to be a disorder of the first pregnancy . In our While we observed no increased risk of gestational diabetes in association with noise exposure in the full cohort, we observed strong risks among the subsample of first pregnancies. Gestational diabetes recurrence is known to be high among those who had it during their first pregnancy, and, therefore, it might seem counterintuitive that risks are not seen in the full cohort including both first and successive pregnancies . We specTo date, the mechanism through which a woman develops HDP and gestational diabetes is unclear. For cardiovascular and metabolic diseases in general, noise is thought to induce a stress response, activating the sympathetic nervous and endocrine systems, leading to increased blood pressure, vascular damage, and metabolic disorders through oxidative stress and increased inflammation . BecauseWith this mechanism in mind, one would expect to find the greatest risk at levels >85 dB(A), in other words, increasing levels of noise leading to an increased stress response; however, in our data this was not the case. One explanation could be that at these levels, workers are required to use hearing protection, which would protect mothers from the any adverse effects of noise. At the 80–85 dB(A) levels, workers are required to have access to, but do not necessarily have to use, hearing protection. One study, however, found no decrease in the risk of gestational hypertension among women who self-reported use of hearing protection . RegardlThis study has some limitations. Regarding exposure ascertainment, the noise levels provided by the JEM are not measured on an individual level, and thus misclassification can occur. The JEM were developed for general use based on measurements on both male and female workers and expert knowledge, without consideration for any particular outcome. Therefore, any misclassification is most likely non-differential and, as such, would only attenuate the risk reported. We did not have information on use of hearing protection. As mentioned, it is possible that the lower risk and lack of significant findings in the highest exposure category is due to use of hearing protection, but we could not test this hypothesis. Similarly, we did not have information on task reassignment. We did have information on pregnancy leave; however, because diagnoses were only given at the end of the pregnancy, we had no way to ascertain that the leave preceded any issues arising from HDP or gestational diabetes. Thus, we could not account for duration of exposure in our analyses. Due to local regulations, task reassignment and leave would most often be associated with physically strenuous work, for which our results are adjusted. Finally, we did not have information on the percentage of time part-time employees work, which can also affect the extent of exposure.On the other hand, this study benefitted from a large sample and a prospective design. Even though HDP and gestational diabetes are the most common conditions of pregnancy, they are still relatively rare in the population. At the same time, the multifactorial nature of these disorders indicates there is likely no one major causative factor. Therefore, only a nationwide register this large can capture the number of exposed cases needed to detect the magnitude of differences expected. Additionally, because the data covers approximately 99% of all births in Sweden, results are generalizable to all pregnant women in Sweden. Another advantage is that data from this national, well-established register were collected as part of routine maternal care by healthcare professionals with nearly complete coverage of all pregnancies; therefore, data are more likely to be accurate and consistent for the entire sample. In this study, we were also able to control for several other exposures that confound the association between noise and pregnancy disorders via other JEM. These JEM were developed by experts in their respective fields and are based on the Swedish workforce, which should reduce misclassification at the occupational groups level.In conclusion, this study indicates that there is an increased risk in HDP associated with exposure to noise at levels >80 dB(A). There is also an increased risk for gestational diabetes associated with these levels for first-time pregnant women who work full-time. Additional studies are needed to confirm this association, as well as investigate whether hearing protection can mitigate these risks.


To evaluate the efficacy of bevacizumab a monoclonal, antivascular endothelial growth factor antibody in combination with cytotoxic chemotherapy in Turkish patients with recurrent and metastatic cervical cancer.Data of 64 patients with metastatic or recurrent cervical cancer, receiving bevacizumab with first-line cisplatin or carboplatin and paclitaxel chemotherapy between 2013 and 2017 were retrospectively evaluated.The mean age of the patients was 49 years , the median follow-up time was 12 months , the median progression-free survival (PFS) was eight months, and the median overall survival (OS) was 23 months. All 64 patients received a median of 6  bevacizumab and 6  chemotherapy cycles. The chemotherapy regimens used with bevacizumab were cisplatin and paclitaxel in 31 (48%) and carboplatin and paclitaxel in 33 (52%) patients. The survival in patients treated with bevacizumab and cisplatin plus paclitaxel was better-particularly in patients with no previous cisplatin-based radiosensitizer therapy-than those treated with carboplatin, paclitaxel, and bevacizumab (p=0.023). The bevacizumab dose was 7.5 mg/kg in 30 patients (47%) and 15 mg/kg in 34 patients (53%) every 21 days. No significant difference was reported in the OS and the PFS between the two groups. While the most common all-grades adverse events were nausea, neutropenia, anemia, and peripheral sensory neuropathy, the most common grade ≥3 adverse events were neutropenia, anemia, and peripheral sensory neuropathy.Adding bevacizumab to platinum and paclitaxel chemotherapy in a case of metastatic or recurrent cervical cancer is an effective and tolerable treatment for Turkish patients. PRECIS: Adding bevacizumab to chemotherapy in advanced stage cervical cancer is an effective and tolerable treatment for Turkish patients.. In 2012, cervical cancer accounted for an estimated 528,000 new cancer cases and for 266,000 deaths worldwide. However, it can be prevented and usually cured if detected early. The therapeutic paradigms in the primary management of cervical cancer are well-established, that is, early lesions are usually treated surgically and locally advanced lesions are managed with concurrent cisplatin chemotherapy and pelvic radiation,5. When women with metastatic disease (stage IV B) are treated with systemic cisplatin combination regimens, the median survival is only 9-10 months,6,7.Cervical cancer is the third most commonly diagnosed gynecologic malignancy and the third most common cause of death among all gynecologic cancers in the United States,9 and this has been supported by a Phase 3 trial in which the survival of the women with metastatic cervical cancer increased when first-line chemotherapy was added. The current retrospective study was conducted to assess the tolerability, safety, and activity of bevacizumab-containing therapy in Turkish patients with advanced cervical cancer.Vascular endothelial growth factor (VEGF) signaling is an important target for cancer therapy because of its role in tumor angiogenesis and its potential role in tumor cell survival. Bevacizumab acts as a monoclonal antibody against VEGFData of 64 patients with radiologically confirmed stage IVB, persistent or recurrent cervical cancer diagnosis, and undergoing platinum (cisplatin or carboplatin) plus paclitaxel and bevacizumab treatment between 2013 and 2017 were retrospectively evaluated.The patients were divided into two categories: those who had previously received chemoradiotherapy (including cisplatin) and those who had not. The patients were classified according to the bevacizumab dose and chemotherapy regimen in which bevacizumab was added to cisplatin or carboplatin, and the metastatic region of the patients was lymphatic or visceral.Ethical approval was received from the Kartal Dr. Lütfi Kırdar Training and Research Hospital, Non-interventional Clinical Research Ethics Committee . No patient consent was required.The overall survival (OS) and the progression-free survival (PFS) were calculated using the Kaplan-Meier method starting from the first day of the chemotherapy. Data were expressed as mean, standard deviation, median lowest value, median highest value, frequency, and rates. All p-values were 2-sided, and p=0.05 was considered as statistically significant. Data were analyzed using the SPSS software version 22.The mean age of the 64 patients whose data were included in this study was 49 years range, 28-68) , and the8-68 , anThe recommended bevacizumab dose was 7.5 mg/kg in 30 patients (47%) and 15 mg/kg in 34 patients (53%) every 21 days. No significant difference was observed in the OS and the PFS between the two groups. While 17 patients (26%) had metastases at the time of diagnosis and the median survival for them was 10 months, 47 patients (74%) had recurrent disease with the median survival of 24 months (p=0.14). The metastatic regions were visceral organs in 25 (39%) and lymphatic region in 39 patients (61%). The median survival was 10 months in patients with visceral and 51 months in those with lymphatic metastases (p=0.10).While the most common all-grade adverse events were nausea, neutropenia, anemia, and peripheral sensory neuropathy, the most common grade ≥3 adverse events were neutropenia, anemia, and peripheral sensory neuropathy. Fistula and toxic death were not observed .. Based on the results of the GOG204 and JCOG0505 studies, paclitaxel/cisplatin and paclitaxel/carboplatin are considered to be the recommended treatments for patients with recurrent cervical cancer,13. VEGF plays a key role in the angiogenesis of the tumor, and although it is directly related to the extent of the disease, it is inversely related to the survival. Bevacizumab is an antibody that recognizes and neutralizes all major isoforms of VEGF, inhibits endothelial cell proliferation, and inhibits receptor binding and vascular formation.This retrospective study was conducted with an aim to assess the tolerability, safety, and activity of bevacizumab-containing therapy in Turkish patients with advanced cervical cancer. Although, chemotherapy remains the standard treatment for patients with stage IVB, persistent or recurrent cervical cancer, no long-term disease control is achieved through this treatmentIn this study, the median PFS was reported as 8 months and the median OS as 23 months, and the overall response rate was 78%. About 39% of the patients had previously received a platinum-based radiosensitizer therapy. The survival with cisplatin, paclitaxel, and bevacizumab treatment was better in patients without a previous cisplatin-based radiosensitizer therapy than those who were treated with carboplatin plus paclitaxel and bevacizumab (p=0.023). No difference was observed in the survival between the two regimens in patients who had previously received cisplatin (p=0.556). Also, there was no significant difference in the OS and the PFS between patients with bevacizumab doses of 7.5 mg/kg (n=30) and patients with 15 mg/kg (n=34). The survival was 10 months in patients with systemic and 51 months in patients with lymphatic metastases. Although there was a numeric difference between the two groups, there was no statistical significance. This may be due to the insufficient number of patients (p=0.10). There was no toxic death due to treatment, no patient developed fistula, and grade ≥3 and higher side effects were in-line with the literature. in 2006 retrospectively evaluated 6 patients who had previously received median 3 systemic treatments. The authors reported that patients receiving bevacizumab with chemotherapy had a response rate of 67% and a median time of 4.3 months to tumor progression. Therefore, in this small cohort study, bevacizumab was well-tolerated.The efficacy of bevacizumab has been seen in a wide range of solid tumor types, including in case of advanced cervical cancer. Wright et al. in 2009, the efficacy of bevacizumab alone in patients receiving first- or second lines of systemic therapy for recurrent disease was investigated. The PFS in the study was reported as 3.4 months and the OS as 7.29 months. It was therefore shown to be an effective and well-tolerated treatment in the second- and third lines of therapy.In the first Phase 2 study conducted by Monk et al. conducted Phase 2 and 3 studies. In this study, 49 local advanced (stage IB-IIIB) patients were treated with chemoradiotherapy and bevacizumab 10 mg/kg every 2 weeks. Researchers concluded that the addition of bevacizumab in chemoradiotherapy is a feasible and safe treatment.Later, in 2011, Schefter et al..In Phase 3 trial, the investigators randomly assigned 452 patients to chemotherapy with or without bevacizumab at a dose of 15 mg/kg. Chemotherapy consisted of cisplatin plus paclitaxel or topotecan, and paclitaxel. Topotecan/paclitaxel was not superior to cisplatin/paclitaxel. Bevacizumab, as compared with chemotherapy alone, was associated with an increased incidence of grade 2 or higher hypertension, grade 3 or higher thromboembolic event, and grade 3 or higher gastrointestinal fistulas. The addition of bevacizumab to a combination chemotherapy in patients with recurrent, persistent, or metastatic cervical cancer was associated with an improvement of 3.7 months in median OS showed that cisplatin plus paclitaxel and bevacizumab treatment are effective, tolerable, and safe in Japanese patients with a small single-arm study (seven patients).Further, in 2016, Sugiyama et al.Although, owing to cytological screening and DNA testing for high-risk human papilloma virus types, the rate of cervical cancer has significantly reduced in developed countries, it, however, remains a major problem in undeveloped countries. If diagnosed at an early stage, patients can be cured by surgery, radiotherapy, and chemoradiotherapy treatments. However, in patients with recurrent metastatic disease or after platinum therapy, survival is poor with systemic treatments, and most cases cannot be cured. The newly targeted therapies and immunotherapy trials in advanced-stage patients continue to produce better survival outcomes and cure. Preventive vaccination and early detection of the disease are very important factors in survival. There is no study in the literature that compares carboplatin-paclitaxel and bevacizumab treatment with cisplatin-paclitaxel and bevacizumab treatment in patients with cervical cancer, and there is no study comparing the dose of bevacizumab 7.5 mg/kg with a dose of 15 mg/kg. In this retrospective study, we aimed to report our experience with such subjects and the efficacy and safety data of bevacizumab in our patient population.Being a retrospective study, covering small number of patients and short follow-up time are the limitations of the study.The introduction of bevacizumab has been one of the most important recent advances for patients with advanced-stage cervical cancer. It is an effective and tolerable treatment for Turkish population with metastatic or recurrent cervical cancer.


Aminoglycosides (AMGs) have been extensively used to treat infectious diseases caused by Gram-negative bacteria in livestock and humans. A selective and sensitive colorimetric probe for the determination of streptomycin and kanamycin was proposed based on chlortetracycline-coated silver nanoparticles (AgNPs–CTC) as the sensing element. Almost all of the tested aminoglycoside antibiotics can rapidly induce the aggregation of AgNPs, along with a color change from yellow to orange/red. The selective detection of aminoglycoside antibiotics, including tobramycin, streptomycin, amikacin, gentamicin, neomycin, and kanamycin, with other types of antibiotics, can be achieved by ultraviolet (UV) spectroscopy. This developed colorimetric assay has ability to detect various AMGs using in-depth surface plasmon resonance (SPR) studies. With this determination of streptomycin and kanamycin was achieved at the picomolar level (pM) by using a UV–visible spectrophotometer. Under aqueous conditions, the linear range of the colorimetric sensor for streptomycin and kanamycin was 1000–1,1000 and 120–480 pM, respectively. The corresponding limit of detection was 2000 pM and 120 pM, respectively. Thus, the validated dual colorimetric and ratiometric method can find various analytical applications for the ultrasensitive and rapid detection of AMG antibiotics in water samples. Aminoglycosides (AMGs) are RNA-binding antibiotics sharing a common core structure named the streptamine ring. The mechanism of action involves binding to bacterial ribosomes, a vital role in the formation of nonsense peptides, finally leading to bacterial cell death . TherefoTest kits and other laboratory-based methods used for the detection of antibiotics are mainly based on microbial inhibition assays. Most of them, however, either cannot detect AMGs or have a much higher limit of detection (LOD) relative to the maximum residue limit . ConventGold nanoparticles (AuNPs) have been used extensively for colorimetric detection of both inorganic and molecular targets in several settings, including water and food safety screening ,8,9. The3), tetracycline, ampicillin, chlortetracycline, streptomycin, and penicillin G were procured from Sigma–Aldrich .Tobramycin, doxycycline, and neomycin were purchased from Alfa Aesar . Oxytetracycline, amikacin, penicillin V, and metacycline were purchased from Cayman . Kanamycin, erythromycin, gentamycin, azithromycin, and clarithromycin were obtained from Tokyo Chemical Industry Co., Ltd . Silver nitrate  under different temperature conditions  was performed to improve the AgNPs synthesis. The colorless reaction mixture rapidly turned brown within a few minutes, showing a well-defined SPR band centered at 400 nm. Then, 0.2 mL aliquots of AgNPs collected under different temperature conditions were diluted to 0.8 mL with nanopure water. At 60 min after the AgNPs synthesis, the ultraviolet–visible (UV–vis) spectra of the samples were recorded in the range 300–800 nm using a spectrophotometer (Optizen 2120) equipped with an automatic rotary type 8-cell holder and 1 cm path length quartz cuvettes. The average of each triplicate measurement (n = 3) was used to observe the absorbance intensity at 400 nm and plotted against the temperature conditions to monitor the productivity. High-resolution transmission electron microscopy (HR-TEM) was performed on a Jeol JSM-2100F  to determine the size distribution and shape of the AgNPs. AgNPs were deposited onto glass and dried before analysis by X-ray photoelectron spectroscopy . A monochromatic Al Kα as an X-ray source was used to obtain the XPS data.Synthesis of AgNPs–CTC was performed with minor modification of the previously reported method . BrieflyThe solution of AgNPs was then purified by ultracentrifugation at 12,000 rpm for 15 min, redispersed in water to remove the excess organic groups from the surface of the AgNPs, and stored at room temperature. However, no color change was observed. The color of the reaction mixture remained yellow, suggesting the successful CTC conjugation and surface functionalization . The lacThe selectivity of any analytical probe is important and investigated by noticing the change in color and red-shifts in the UV–vis spectra. The AMG antibiotics, including streptomycin, kanamycin, tobramycin, neomycin, amikacin, and gentamycin, were prepared by treating 0.1 mL of 1 µM stock solutions with the stock solution of AgNPs (50 μL) in 0.85 mL of water. For the application of the AgNPs probe as a colorimetric indicator specific to the AMGs, other antibiotics, including penicillin G, penicillin V, ampicillin, erythromycin, clarithromycin, azithromycin, tetracycline, chlortetracycline, oxytetracycline, metacycline, and doxycycline were tested under identical conditions. The UV–vis spectra and color of the AgNPs probe were recorded for all tested AMGs and other antibiotic samples after 20 min of reaction at ambient temperature (22–24 °C).n = 3) using a 30 μL AgNPs probe solution. The UV-vis spectra and ratiometric absorbance (A540/A400) were recorded for 20 min of treatment at ambient temperature. The LOD of the prepared AgNPs probe for both streptomycin and kanamycin was calculated, and the average value reported.Quantification of the AMGs, both streptomycin  and kanamycin (120–480 pM) was performed in triplicate (n = 3). Streptomycin  and kanamycin  were then added, and the solutions mixed well. The AgNPs probe was allowed to interact with the target antibiotics for another 20 min at ambient temperature. The absorbance intensities of the dilute NaOH-treated AgNPs probe were again measured at 540 nm.The efficiency of the AgNPs probe was studied as a function of the ionic strength  using 30  μL aliquots of the AgNPs stock solution The AgNPs probe and NaOH solutions were thoroughly mixed in UV–vis cuvettes. After incubation for 5 min, the absorbance intensities were measured at 540 nm . The absorbance intensities at 400 and 540 nm were remeasured at 2 min intervals for up to 18 min.+ ions. The addition of AgNO3  to the preheated basic CTC solution was performed under different temperature conditions . The resultant formation of AgNPs was monitored for 60 min. The UV–vis spectra and absorbance results  of CTC molecule functionalities. The CTC molecule is complex and shows different tautomeric forms, so it is difficult to determine the structure of CTC on the surface of NPs. However, according to the N1s spectra, CTC may conjugate to the AgNPs surface by an amide linkage because Ag+ ions have a high affinity for amide group-containing materials [The AgNPs had an average size of about 14 nm and were monodispersed with mixed shapes but a predominance of spherical NPs, as illustrated in the representative electron micrograph c. It wase values ,19. As saterials ,21.To investigate the selectivity of the AgNPs probe, potentially interfering competitive antibiotics, such as streptomycin, kanamycin, tobramycin, neomycin, amikacin, and gentamycin of the AMG class, and other antibiotics, including penicillin G, penicillin V, ampicillin, erythromycin, clarithromycin, azithromycin, tetracycline, chlortetracycline, oxytetracycline, metacycline, and doxycycline, were performed under identical conditions. The corresponding visual appearance of each AgNPs probe sample is shown in the inset of Although effective in the treatment of various infectious diseases, AMG antibiotics have severe side effects, which limit their clinical use ,23,24. TMost of the antibiotics that come under the class of AMGs carry a positive charge. Remarkably, however, not all of them cause aggregation of the AgNPs probe identically. When the ability of six different AMGs to cause the red-shift of the AgNPs probe was investigated under identical conditions, almost all the AMGs were distinguishable from the control sample. As can be seen in n = 3) was performed using a relatively small sample volume (1 mL) within 20 min. It is evident in 540/A400 absorbance wavelengths was calculated to provide a linear concentration plot between 1000 and 11,000 pM streptomycin (2) of 0.964, for streptomycin. Thus, this proposed probe is a highly sensitive and colorimetric detection system for all kinds of AMG antibiotics at concentrations (picomolar) below the safe limit set for drinking water. The LOD of the AgNPs method is comparable to or lower than those of the reported methods for detecting streptomycin  was investigated. This study is also vital to determine the effect of NaCl for CTC coated AgNP probe against salt-induced aggregation. Aggregation of the AgNPs occurred with the increasing addition of NaCl . SubsequFrom The time-dependent absorbance intensity  was alsoAt all tested concentrations of kanamycin, a stable response for the AgNPs–CTC was observed after an initial increase to the maximum. Such findings indicate the possibility of rapid detection of AMG antibiotics in the water samples within the time frame of 5 to 20 min. In comparison, the colorimetric results, which were highly reproducible and reliable, were visible for a longer period (data not shown). The color of the reaction mixture remained stable for a time frame of minutes to hours, without the formation of agglomeration or precipitation.Herein, we have shown that SPR can be used to analyze various AMG antibiotics. For the first time, a CTC-coated AgNPs probe was reported for the rapid colorimetric detection of AMG antibiotics in an aqueous system. The organic ligand-induced aggregation of the AgNPs–CTC probe was found to be a sensitive and cost-effective ratiometric assay for AMG antibiotics. The AMG antibiotics serve not only as target analytes but also as molecular linkers for electrostatic coupling with the AgNPs–CTC probe, giving rise to a noticeable color change from yellow to red that can also be detected by UV–vis spectroscopy. Analysis of streptomycin and kanamycin in water samples was demonstrated, with excellent picomolar-level sensitivity. Thus, this validated ratiometric probe can find analytical applications in the ultrasensitive detection of AMG antibiotics.


In bench press exercises, time to PV was lower in WU compared with WU40  and total work was higher . The results showed that force outputs were mainly optimized by WU80 in squat training and by WU in bench press training. Moreover, warming-up with few repetitions and low loads is not enough to optimize squat and bench press performances.The current study aims to verify the effects of three specific warm-ups on squat and bench press resistance training. Forty resistance-trained males (19–30 years) performed 3 × 6 repetitions with 80% of maximal dynamic strength (designated as training load) after one of the following warm-ups (48 h between): (i) 2 × 6 repetitions with 40% and 80% of the training load (WU), (ii) 6 × 80% of training load (WU80), or (iii) 6 × 40% of the training load (WU40). Mean propulsive velocity (MPV), velocity loss (VL), peak velocity (PV), time to achieve PV, power, work, heart rates, and ratings of perceived exertion were analyzed. In squat exercises, higher MPV were found in WU80 compared with WU40 (2nd set: 0.69 ± 0.09 vs. 0.67 ± 0.06 m.s Warm-up has been identified as essential to maximizing the athlete’s performance in different sports and physical activities . The actPrevious research has found an increase of 3% of upper body power during medicine ball throws after specific warm-ups in bench press exercises with the maximal external load that each subject can move properly for 5 repetitions, contrary to higher or lower loads . The numStrength performance can be improved using a specific warm-up ,14; neveKnowing the importance of warm-ups for strength optimization and the need to avoid the effects of muscle fatigue, it is important to observe the effects and influence of using different specific warm-up designs to increase the efficiency of resistance training exercise performance. For instance, during resistance training, an appropriate warm-up would increase force development against the same training external load, improving the training efficiency, training intensity, and, therefore, long-term adaptations to develop higher force values and/or higher velocities ,21,22,23The current study tried to further understand the importance of warm-up intensities for resistance training exercises performance and then to realize the optimal warm-up load that should be used pretraining. The effects of the warm-up load in resistance training performance were evaluated by analyzing the bar movement velocity using a recent validated device ,24,25,26A crossover research design was used to analyze the effects of a warm-up on mechanical responses, mean propulsive velocity (MPV), propulsive mechanical power (MPP), and physiological (heart rate) and psychophysiological variables (ratings of perceived exertion: RPE). After the adaptation session, four evaluation sessions were performed on four different days (48 h in between). The first evaluation was to determine the 1RM load of each participant in squat or bench press exercise. Then, each participant performed resistance training (3 sets of 6 repetitions at 80% 1RM loads) in three subsequent sessions. These resistance exercises were chosen because of their relevance in resistance training, thus commonly used by conditioning specialists and coaches for sports training. Moreover, the intensity of 80% 1RM is widely used in traditional resistance training, and it is in the optimal range of relative intensities (30–80% 1RM) that have been suggested to improve long term muscular performance ,28,29. TForty men aged 19–30 years old volunteered to participate in the current study. From these, 14 men  were evaluated using squat exercises, and 26 men  were assessed using bench-press exercises. Subjects were physically active, engaged in physical activity regularly, with an experience of resistance training for the last two years. Everyone was asked to report any previous illness, injury, or other physical issues that would hinder their performance. The eligibility criteria were being healthy and injury-free, aged between 18 and 35 years old, and having previous experience with back-squat and bench-press exercises. Criteria of exclusion were the evidence of an orthopedic or medical problem or another self-reported issue that would endanger their health. The participants were informed about the study procedures and written informed consent was signed. The investigation was conducted in accordance with the Declaration of Helsinki and was approved by the University of Beira Interior Research Ethics Committee .The first session was used for body composition evaluation  and adaptation to testing conditions, where the subjects performed some repetitions with lower loads in bench press or squat exercises, with some orientation about the correct technique. The second session was used to determine the individual load–velocity relationships and estimated 1RM strength in the squat or bench press exercises using a progressive loading test. Then, the last three sessions were used to evaluate performance during a resistance training session with different specific intensities of warm-ups. All procedures were performed using a Smith machine , similar to other studies, e.g., ,25,26. TIn the squat exercise, each subject started from the upright position with the knees and hips fully extended, stance approximately shoulder-width apart, and the barbell resting across the back at the level of the acromion. The eccentric phase was performed in a continuous motion until the top of the thighs was below the horizontal plane, and the concentric phase was made at the maximum velocity to the initial position ,25. Trai−1 in the bench press and 0.6 ms−1 in the squat was attained, corresponding to 85% 1RM in both [To determine individual 1RM loads, the initial load was set at 20 and 30 kg for all participants in the bench press and squat exercises, respectively, and was gradually increased by 10 kg increments. The test ended for each participant when the concentric MPV of 0.4 ms in both ,25. Thre in both ,29. The  in both .Three different warm-up conditions were randomly implemented. WU comprised 6 repetitions with 40% of the training load, followed by 6 repetitions with 80% of the training load . The warm-up with high intensity included 1 set of 6 repetitions with 80% of the training load (WU80), and the warm-up with low intensity comprised 1 set of 6 repetitions with 40% of the training load (WU40). The effect of performing warm-ups was assessed during a bench press or squat resistance training session performed 3 min after the warm-up. Resistance training consisted of 3 sets of 6 repetitions with 80% 1RM load, with 3 min of interset recovery. The number of sets and loads intensities were selected because of their common use in resistance training in diverse competitive sports and their effects on muscular development and performance improvement ,31. DuriAll velocity measures identified in this study corresponded to the propulsive phase of each repetition ,21,24. TThe heart rate values were assessed at rest (baseline), 1 min after the warm-up, and immediately after training (1 min). Additionally, the rating of perceived exertion (RPE) values was recorded using a 16-points Borg scale (6–20 rates)  immediatt-tests were run to additional investigate the effect of each condition. The effect size was calculated to estimate the variance between conditions , and Cohen’s dz (ES) for within-subject comparisons were calculated using the Excel spreadsheet by Lakens [p2), the cut-off values were interpreted as 0.01 for small, 0.09 for moderate, and 0.25 for large, and ES values of 0.20, 0.60, 1.20, and 2.00 were considered small, moderate, large, and very large magnitudes, respectively [Standard statistical procedures were selected to calculate means, standard deviations (SDs), and 95% confidence intervals. The normality of all distributions was verified with the Shapiro–Wilk test, and parametric statistical analysis was adopted. The effect of the warm-up procedures was analyzed by an ANOVA for repeated measures, with sphericity checked using Mauchly’s test. Posthoc paired y Lakens . For parectively . Statistp = 0.29, ηp2 = 0.09). However, MPV showed to be moderately affected by warm-up conditions in the second  and third sets . In these sets, the participants were able to perform the squat exercises at higher MPV values after WU80, comparing to WU40. The minimal value of MPV in WU40 showed lower values when compared with the other warm-ups during the second set . No significant differences were found between conditions in VL during the first , second  and third sets . Curiously, the values were moderately higher in WU40 during the second set. In The mean values, differences, and ES for MPVs in the first, second, and third sets in the squat exercise are presented in p = 0.06, ηp2 = 0.19).The mean values, differences, and ES for PV and time spent to achieve PV in the first, second, and third sets in the squat exercise are presented in p = 0.46, ηp2 = 0.06), second , and third sets , moderate effects were found with lower values in the WU40 condition. Large effects were found in the minimal values of MPP in the second set , with higher values recorded in WU80 when progressively compared with WU40. No differences were found in the work produced during squat resistance training .p = 0.10, ηp2 = 0.09) and second sets  after WU warm-up when compared with WU40  and second sets . The total work also presented significant results between these two conditions p = 0.19, np2 = 0.06) and in the squat exercise . In the perceived effort after the bench press training, no differences were found between conditions , and no differences existed regarding the squat training .Immediately after performing the three sets of resistance training, no differences were found in heart rate values in the bench press exercise . Despite these limitations, the results are interesting for all coaches, athletes, and researchers. Most of the studies investigating the effects of warm-ups on muscle force output generally focus on maximal dynamic strength and the maximal number of repetitions, disregarding the confounding effect of the procedures . Linear position technology, commonly used to measure force outputs and monitor athletic performance, allowed us to measure and analyze the changes in resistance training caused by different specific warm-ups.The current study took a novel approach to warm-up research by examining the effects of warm-ups on resistance training using some recent procedures to measure performance. In summary, an increase in force outputs during squat training was found after a specific warm-up, comprising only six repetitions with 80% of the training load. This was clear during the second and third training sets. In the bench press exercise, a progressive-intensity warm-up of two sets of 6 repetitions, with 40% and 80% of training load, was the most effective, resulting in less time to attain maximal velocities and augmented work. In general, the warm-up with 40% of the training load was the one with worse performances in both exercises.Some practical applications can be drawn. Despite some different responses for each exercise, progressive-intensity specific warm-up and WU80 should be used to maximize performance during resistance training. Specifically, our outcomes allowed us to disclose that higher intensities  and only a few repetitions  should be used during specific warm-ups to optimize squat training. However, the bench press exercise should be preceded by more repetitions  with progressive loads . The present findings can be applied before bench press and squat resistance training but also when assessing maximal dynamic strength. Knowing that the movement velocity can be used to determine maximal dynamic strength and to monitor resistance training performance, our outcomes can provide new insights and recommendations for sports professionals and researchers to improve training efficiency and optimize performance.