COVID-19 caused by the SARS-CoV-2 infection may result in various disease symptoms and severity, ranging from asymptomatic, through mildly symptomatic, up to very severe and even fatal cases. Although environmental, clinical, and social factors play important roles in both susceptibility to the SARS-CoV-2 infection and progress of COVID-19 disease, it is becoming evident that both pathogen and host genetic factors are important too. In this study, we report findings from whole-exome sequencing (WES) of 27 individuals who died due to COVID-19, especially focusing on frequencies of DNA variants in genes previously associated with the SARS-CoV-2 infection and the severity of COVID-19.We selected the risk DNA variants/alleles or target genes using four different approaches: 1) aggregated GWAS results from the GWAS Catalog; 2) selected publications from PubMed; 3) the aggregated results of the Host Genetics Initiative database; and 4) a commercial DNA variant annotation/interpretation tool providing its own knowledgebase. We divided these variants/genes into those reported to influence the susceptibility to the SARS-CoV-2 infection and those influencing the severity of COVID-19. Based on the above, we compared the frequencies of alleles found in the fatal COVID-19 cases to the frequencies identified in two population control datasets (non-Finnish European population from the gnomAD database and genomic frequencies specific for the Slovak population from our own database). When compared to both control population datasets, our analyses indicated a trend of higher frequencies of severe COVID-19 associated risk alleles among fatal COVID-19 cases. This trend reached statistical significance specifically when using the HGI-derived variant list. We also analysed other approaches to WES data evaluation, demonstrating its utility as well as limitations.Although our results proved the likely involvement of host genetic factors pointed out by previous studies looking into severity of COVID-19 disease, careful considerations of the molecular-testing strategies and the evaluated genomic positions may have a strong impact on the utility of genomic testing.The online version contains supplementary material available at 10.1186/s12864-022-09084-5. The firThe mortality rate of COVID-19 (ranges between 1–7%) is lower than that of the other two types of coronaviruses, the severe acute respiratory syndrome (SARS-CoV) and the middle east respiratory syndrome (MERS-CoV). However, the rate of human-to-human transmission is higher, as the virus can primarily be transmitted through respiratory droplets and close contact , 6–10. CSARS-CoV-2 infection exhibits varied infectivity and mortality rates in different worldwide populations , 22. An https://www.covid19hg.org/) is currently leading a worldwide public effort to analyse COVID-19 information in millions of individuals in relation with genotype data. It aims to identify genetic variants associated with SARS-CoV-2 infection as well as COVID-19 rate of hospitalization and disease severity. A recent study by Baggen et al. summarizes the wealth of information on proviral SARS-CoV-2 host factors that have been produced by genome-wide functional genetic screens and interactome analyses with further discussion about their roles in cellular processes [In this respect, the COVID-19 Host Genetics Initiative (HGI), 2020  showed evidence of association with COVID-19 in GWAS studies, b.) were located in genes previously reported to be a risk in selected studies by literature search, c.) showed strong association in meta-analyses conducted by the COVID-19 HGI Browser, and d.) were located in genes associated with COVID-19 by QIAGEN Clinical Insight (QCI™) Analyze software. We present a comprehensive comparative assessment of variants identified in these four groups compared to our cohort of genotyped patients. In addition, allele frequencies of identified variants were compared with two types of control groups, worldwide public data of the non-Finnish European population from the gnomAD database (NFE) and our genetic data from Non-Invasive Prenatal Testing (NIPT) in the Slovak population. Despite the growing knowledge about genetic factors associated with the risk of COVID-19, there is a lack of studies that elucidate the multitude of other factors that influence this disease. In this study, we provide different approaches to analysing genetic variants in the WES data from Slovak patients who have died of COVID-19.https://www.ebi.ac.uk/gwas/). Overall, 413 risk variants in a total of 403 genes were identified in the GWAS dataset after entering the keyword “COVID-19” into the GWAS Catalog Browser. After merging all identified variants from GWAS (413 risk variants) with our WES data of the deceased patients, we identified only 3 common variants – the rs11147040 missense variant, the synonymous rs72472161 variant, and the rs8176719 coding sequence variant. Other identified variants belonged to the intron, non-coding, and UTR variants group.The first goal of our analysis was to investigate the variants associated with COVID-19 in GWAS Catalog , 154 (II.), and 437 (III.) common risk variants, respectively. Next, we compared this data with two control groups—the NIPT data from the Slovak population, where we identified 103 (I.), 96 (II.), and 245 (III.) common risk variants and the NFE data  in the I. group. Also, we identified the rs2498800 risk variant  in the III. group after comparison between the NIPT and WES data. Further still, the rs7853989, rs8176741, rs8176743, rs8176746, rs8176747, rs8176749, and rs8176751 risk variants were found in the II. group based on comparison between the WES and NFE data in the overlap of the publications data/WES/NIPT/NFE , we have identified 9, 12, and 10 risk variants with https://app.covid19hg.org/). The total number of COVID-19 missense risk variants found in each HGI group is summarized in Table In this part, we analysed the COVID-19 risk variants identified in meta-analyses conducted by the COVID-19 HGI Browser. We focused only on missense variants from all four genome-wide meta-analysis results available online  as well as between the WES data and the NFE data , both in the B2 group in the HGI groups data/WES/NIPT/NFE overlap  in the comparison between the WES and NIPT data in the A2 group. Equally, 4 risk variants, rs8176747, rs8176746, rs8176743 and rs7853989  were identified in the comparison between the WES and NFE data in the C2 group  for each patient and then applying a biological filter, we selected 4 from the 8 offered COVID-19 filters. Variants are divided into two categories to follow the concept applied in other analyses: the QCI infection group—containing 376 risk variants with the "Mild COVID-19" biological filter and the QCI critical group containing 1738 variants with "Critical COVID-19", "Severe COVID-19" and "COVID-19 related immunodeficiency 74" biological filters. Other filters were excluded from further analyses as it was not possible to clearly determine their classification into infected or critical groups. Subsequently, a number of risk variants in the WES/NFE comparison or in the NIPT/WES/pub comparison or HGI data were contrasted to the QCI analysis of the infection and critical groups. The results of individual comparisons of the number of variants are shown in Table In the present article, we attempted to elaborate on the spectrum of risk variants and genes identified in different ways and their possible relationship to severity and/or mortality of COVID-19. We investigated the frequencies of these variants and evaluated their possible role using a cohort of 27 Slovak patients who have died of COVID-19. In our previous studies, we described the re-use of the data from NIPT for genome-scale population-specific frequency determination of small DNA variants , copy nuOver the past two years, GWAS have offered the opportunity to reveal factors of genetic susceptibility factors to COVID-19 disease and provided insights into the biological basis of SARS-CoV-2 etiology. To date, a large number of risk genetic variants and genes have been identified by the GWAS approach, which has been intimately connected to the COVID-19 susceptibility and severity –35, 48. CDSN (Corneodesmosin) gene and four variants  all located in the ABO gene. Two missense variants, rs8176747 and rs8176746, were found by a comparison between the WES and NFE data in the C2 HGI group and the II. group in the analysis using literature search. Recently, a GWAS found a COVID-19-association signal at locus 9q34.2 coincident with the ABO-blood group  in Italian and Spanish severe COVID-19 patients with respiratory failure [ACE1 (Angiotensin-converting enzyme 1), ACE2 (Angiotensin-converting enzyme 2), and ABO-locus  in COVID-19 prognosis, suspecting inherited genetic predispositions to be predictive of COVID-19 severity [In 2021, a large consortium conducted some highly anticipated studies published last year. The COVID‐19 HGI presented results from three genome‐wide association meta‐analyses of up to 49,562 COVID‐19 patients from 46 studies spanning across 19 countries . Similar failure , 50. Anoseverity .Since 2017, the QIAGEN Clinical Insight (QCI™) Analyze software has been available as an integrated clinical decision support solution for the annotation, interpretation and reporting of NGS data. QCI seeks to extract clinical significance and actionability from sequencing data . After aOur study has several key shortcomings. First, although we used different approaches to analyse the WES data of patients who died of COVID-19, we were able to identify only 10 risk variants with significant difference (after the Bonferroni correction) in allele distribution of the WES data and data of Slovak NIPT and NFE. Next, the sample size of the deceased patients was relatively small. Moreover, data from NIPT are strongly biased towards the healthy population of females. It should also be noted that the total number of analyzed risk variants identified by COVID-19 HGI was also relatively small as we focused only on missense variants. We targeted missense variants to identify as many risk variants as possible since most of the identified variants from HGI belonged to the non-coding category. In GWAS analysis, this proved to be useless for analysis of our WES data.As the COVID-19 pandemic led to a global crisis with a serious global impact, it is crucial to determine how host genetic factors are linked to the clinical outcomes. Therefore, many studies have focused on discovering the influence of host genetic factors on the SARS-CoV-2 infection risk or COVID-19 severity. The main aim of our study was to provide an informative insight into the utility of COVID-19 data from publicly available databases with practical application to WES data from COVID-19 patients. WES is routinely used and is gradually being optimized for the detection of rare and common genetic variants in humans. Genetic stratification of patients with COVID-19 from WES data would be possible, but in clinical practice, this approach requires overcoming several limitations that we also faced in our study. Moreover, WES only analyses exonic variants; therefore, the study may have failed to detect potentially related intronic variants. However, whole-genome sequencing (WGS) is becoming increasingly attractive as an alternative, due to its broader coverage, decreasing cost, and more comprehensive information.The method consists of four main steps, presented in the diagram in Fig. https://www.ebi.ac.uk/gwas/search?query=COVID-19. We also performed a literature search of risk genes and genetic variants associated with infection and severity of COVID-19 published in 2020 and 2021 via the PubMed database (https://pubmed.ncbi.nlm.nih.gov/). More details are available in supplementary material. COVID-19 HGI GWAS meta-analysis release 6 data was downloaded from https://app.covid19hg.org/. Details for each study are provided on the HGI website and in the consortium paper [In the second step, we performed a selection of risk variants and/or genes associated with COVID-19. All risk variants associated with COVID-19 found in the GWAS Catalog were downloaded from um paper .https://gnomad.broadinstitute.org/downloads). The detailed information about the NIPT dataset generated and analysed in this study is fully described in our previous studies [In the third step, we compared each group of identified risk variants with our WES data of the deceased patients and with two groups of controls: NIPT data from the Slovak population and data of NFE  Analyze software was used to analyse the WES data of the deceased patients. Variants were imported as an industry-standard VCF into the QCI-A web interface, which enables data interpretation. We performed variant filtering based on biological context with “COVID-19” as the keyword which included eight filters.Python language libraries scikit-allele were used for analysing genetic variation data and matplotlib and seaborn for the graphic representation. The significance of our findings was evaluated using statistical tests implemented in the Python SciPy package. We used the Mann–Whitney U test to compare risk variants allele frequencies from the deceased COVID-19 patients and the Slovak NIPT data or respectively gnomAD data respectively as controls. In addition, observed allele frequencies for each risk variant in all groups were tested using the Fisher's exact test.Additional file 1: Table S1. Genes selected after literature search from 3 types of publications.Additional file 2: Table S2. All 900 Covid-19 associated risk variants used in our analyses with Fisher's exact test p-values. • /p-value - included in the analysis; sig - significance: * - p-value < 0.05, ** - significant after Bonferroni correction; # - overlap with risk variants from QCI infection group; § - overlap with risk variants from QCI critical group; sN - Slovak NIPT data, W - WES data of Slovak dead patients, N - non-Finnish European gnomAD data.Additional file 3. Methods [ Methods –65.


Breast adenoid cystic carcinoma (AdCC) is a rare subtype of triple negative breast cancer. Two morphologic variants are described, namely classic AdCC (C-AdCC) and solid basaloid (SB-AdCC). Recent studies have shown that the SB-AdCC variant has significantly worse prognosis than C-AdCC. Due to the rarity of SB-AdCC, no standard recommendations are available for its management. Data on the use and benefit of chemotherapy in patients with SB-AdCC are sparse and the response to neoadjuvant chemotherapy has not been reported. We present the clinical and pathologic findings of a patient with SB-AdCC treated with neoadjuvant chemotherapy who achieved a remarkable pathologic response. MYB-NFIB fusion gene, followed by fusion genes involving MYBL1 or MYB gene amplification1. The solid basaloid variant of AdCC (SB-AdCC) is composed of cells with scant cytoplasm, marked nuclear atypia and increased mitotic activity2. In contrast to the classic form of AdCC (C-AdCC), which typically follows a relatively indolent course, the SB-AdCC variant has an aggressive clinical behavior4. Vanishingly rare (there are less than 100 published cases of SB-AdCC), currently there are no standardized recommendations regarding management. In the limited series available, most patients were treated with surgical excision and post-operative radiation but adjuvant chemotherapy was often omitted, similar to the management of C-AdCC7. At present, there is a paucity of data on chemotherapy benefit for SB-AdCC and its response to neoadjuvant treatment is unknown. Here, we describe a case of SB-AdCC treated with neoadjuvant chemotherapy with a near complete pathologic response.Breast adenoid cystic carcinoma (AdCC) is a rare histologic subtype of breast cancer, usually of triple negative phenotype. These tumors account for <1% of all breast cancers, composed of a dual population of epithelial and myoepithelial neoplastic cells, arranged in cribriform, tubular and/or solid growth patterns. AdCCs have been shown to be driven by activation of the MYB pathway, most commonly due to a A 56 years old woman of Ashkenazi Jewish descent with no significant past medical history or family history of breast or ovarian cancer, initially presented to an outside hospital after palpating a right breast mass. The patient underwent bilateral mammogram that revealed a right breast 9 mm spiculated mass with associated calcifications and nipple retraction, corresponding to the sonographic finding of an 8 mm irregular, retroareolar mass which appeared to invade the nipple-areolar complex. Ultrasound of the right axilla showed a suspicious lymph node. An ultrasound guided biopsy of the breast mass yielded an infiltrating moderately differentiated mammary carcinoma with associated intermediate grade intraductal carcinoma, solid and cribriform type. Subsequently, the patient presented to Memorial Sloan Kettering Cancer Center (MSK) for cancer care.MYB translocation was identified by fluorescence in situ hybridization (FISH) utilizing break apart probes. The tumor lacked estrogen receptor (ER), progesterone receptor (PR) and human epidermal growth factor receptor-2 (HER2) expression. The Ki67 proliferation index was approximately 40%  and mastectomy, along with sentinel lymph node biopsy were discussed with the patient. Due to the central location of the tumor and imaging suggesting involvement of areolar skin, a wide excision would also require removal of the nipple areolar complex. In this context, a medical oncology consultation was obtained. Given the triple negative phenotype and biopsy proven positive lymph node status, neoadjuvant chemotherapy was recommended because if it could provide a complete pathologic response in the axilla that would obviate the need for axillary dissection. The patient elected to have an attempt at BCS and thus was given pre-operative dose-dense AC-T (doxorubicin plus cyclophosphamide each two weeks with G-CSF support for four cycles followed by carboplatin each 3 weeks for four cycles plus twelve cycles of weekly paclitaxel), which was well tolerated. Because AdCC is often regarded to be a chemotherapy-resistant subtype of breast cancer the patient was monitored with monthly imaging studies to assess response and ensure there was continued shrinkage of disease. After FDA approval, she was also started on pembrolizumab. However, she developed immune-mediated nephritis after the second dose of pembrolizumab resulting in its discontinuation. An MRI was performed following completion of therapy which showed very faint, residual 7 mm linear enhancement extending towards the nipple, significantly decreased from prior study Fig. . On pre-The resection specimen obtained post-neoadjuvant treatment showed a single focus of residual invasive carcinoma, spanning <0.5 mm microscopically, along with approximately 5 mm of residual in situ carcinoma, present in a fibrotic tumor bed grossly measuring 12 × 7 mm Fig. . The car2, in which they detailed 9 cases collected over a period of 11 years. While none of the patients in their series had evidence of local recurrence or distant metastases despite a follow up period of up to 7.3 years (mean 32 months), the data presented in that study suggested that SB-AdCC had a higher propensity for lymph node metastases than C-AdCC. Subsequent studies confirmed this observation, which lead to the recommendation for routine axillary staging in these patients, and also noted more aggressive clinical behavior8. In the largest series reported to date, Schwartz et al.6 compared the clinicopathologic features and follow-up data of 29 SB-AdCC and 75 C-AdCC from MSK. SB-AdCCs had significantly higher histologic grade, higher rates of lymphovascular invasion and lymph node metastases, and were more likely to recur locally and develop distant metastatic disease. In contrast, C-AdCC had a favorable clinical course, with higher survival rates than invasive breast carcinoma of no special type, and 5- and 10-year disease free survival (DFS) rates greater than 95 and 90%, respectively8. In the cohort described by ref. 6, the surgical approach and use of adjuvant radiotherapy were similar between the variants, however patients with SB-AdCC were significantly more likely to receive adjuvant chemotherapy than C-AdCC . Despite this, DFS was significantly reduced in patients with SB-AdCC, with a median DFS of 46.5 months for SB-AdCC versus 151.8 months in those with C-AdCC. While the optimal chemotherapy regimen and benefit of chemotherapy for SB-AdCC are unknown, the poor outcomes of the solid and basaloid variant suggest systemic therapy may be warranted. We present a case of SB-AdCC treated with platinum and anthracycline containing neoadjuvant chemotherapy which achieved an excellent pathologic response, showing complete resolution of the patient’s axillary disease and less than 0.5 mm of residual invasive carcinoma.The mammary SB-AdCC variant was first described in a report by Shin and Rosen9 performed a systematic review of 34 studies, including prospective and retrospective data, examining the systemic treatment of metastatic or locally recurrent AdCC, all but one of which included only tumors of salivary gland origin. In the trials which looked at single agent cytotoxic therapy, the observation of stable disease, noted in 58% (64/111) of patients, was more common than objective major responses which were seen in only 13% (18/141) of patients. Overall, in trials utilizing combination therapy, 25% of patients responded, most commonly to regimens composed of cisplatin, doxorubicin and cyclophosphamide (CAP)9. These results should be interpreted with caution, though, as none of these multi-agent trials enrolled more than 12 patients. The authors also found discrepant response rates amongst reports, with higher rates observed in retrospective reviews versus clinical trials and ultimately conclude further study is needed to develop a standardized management approach. The patient reported here had a remarkable response, providing further support to the notion that chemotherapy may be effective in at least a subset of SB-AdCCs.Published data on the effect of chemotherapy in patients with SB-AdCC is based on small trials and more frequently involve AdCC arising in organs other than the breast. Laurie et al.MYB-NFIB fusion gene is identified in up to 86% of C-AdCC and has been confirmed to be present in the solid-basaloid variant as well, however it is seen much less frequently, reported in only 12.5% of cases in one series10. Additionally, MYB expression by IHC has been observed in up to 63% of other basal-like, triple negative breast cancers and is not pathognomonic for SB-AdCC11. We do not recommend using FISH or IHC testing for MYB unless there are morphologic features strongly raising the differential for SB-AdCC, to avoid overdiagnosis. The most definitive feature pointing to the diagnosis is the identification of areas showing C-AdCC features, particularly identifying the dual cell population and pseudolumina filled with basement membrane material. In the core biopsy presented here the tumor showed focal “cribriforming” areas along with typical basaloid cytologic features suggesting the diagnosis. Another feature of SB-AdCC is the characteristic nested growth pattern embedded in a markedly desmoplastic stroma which contrasts with the typically solid, dense mass seen in poorly differentiated triple negative tumors of no special type.Additionally, novel drug therapies targeting specific molecular alterations may prove to be efficacious in the rare SB-AdCC. The translocation t leading to the MYB gene, recent analysis has revealed that ATR gene expression is significantly upregulated in MYB overexpressing cells12. Andersson et al.12 observed a dose dependent decrease in proliferation and increase in apoptosis following inhibition of ATR, a gene involved in DNA damage repair, and in vivo experimentation demonstrated significant tumor suppression and, in one case, tumor regression, supporting the development of therapeutic targets to ATR in the treatment of C-AdCC and possibly a subset of SB-AdCC. Furthermore, it has been reported that the mutation rate of NOTCH genes is significantly higher in SB-AdCC compared to C-AdCC3. Clinical trials involving Notch signaling inhibitors are currently ongoing and may show usefulness in this variant3.While there are currently no drugs available to target the In conclusion, SB-AdCC is a rare subtype of AdCC. While scant data on optimal management are available, we demonstrate that neoadjuvant chemotherapy may induce an excellent response. Our case suggests that combination chemotherapy has a role in treatment. If ongoing studies are successful, additional targeted therapies could provide desirable outcomes in this more aggressive subtype.Written informed consent was obtained from the patient to publish this case report and associated clinical details. Clinical and radiologic data for this case were obtained from the electronic medical record. IHC was performed according to manufacturer’s manual. FISH was done using DNA breakapart signal analysis.


Luminal areas of seven conducting airways were analyzed, including trachea, left and right main bronchus, intermediate bronchus, left and right upper lobe, and left lower lobe. For the central conducting airways, luminal area was ~ 15% greater patients with COVID-19 compared to matched controls (p < 0.05). Among patients with COVID-19, there were generally no differences in the luminal areas of the conducting airways between hospitalized patients compared to patients who did not require COVID-19-related hospitalization. Our findings suggest that males and females with COVID-19 have pathologically larger conducting airway luminal areas than healthy, sex- and height-matched controls.Respiratory epithelium in the conducting airways of the human body is one of the primary targets of SARS-CoV-2 infection, however, there is a paucity of studies describing the association between COVID-19 and physical characteristics of the conducting airways. To better understand the pathophysiology of COVID-19 on the size of larger conducting airways, we determined the luminal area of the central airways in patients with a history of COVID-19 compared to a height-matched cohort of controls using a case–control study design. Using three-dimensional reconstruction from low-dose high-resolution computed tomography, we retrospectively assessed airway luminal cross-sectional area in 114 patients with COVID-19  and 114 healthy, sex- and height-matched controls . People with a history of smoking, cardiopulmonary disease, or a body mass index greater than 40 kg·m Cells of the conducting airways and gas exchange surfaces of the lungs share a common receptor for SARS-CoV-2, and much of the COVID-related symptomatology manifests in the conducting airways and lungs4.Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the causative agent of novel coronavirus disease 2019 (COVID-19), is primarily transmitted through respiratory droplets, and attaches to ciliated epithelium in the conducting airways of the respiratory tree6 and chest computed tomography (CT) have helped guide diagnosis and treatment of COVID-198, but there is a paucity of studies describing the association between COVID-19 and physical characteristics of the conducting airways. Previous studies provide evidence of an increase in diameter of the trachea proportional to severity of COVID-19 pneumonia9, suggesting that severe inflammation is associated with edema in the trachea and an increase in diameter of the trachea among patients with COVID-1910. However, previous studies are limited by focusing on luminal area of the trachea or by not including a comparator group without COVID-1910.Both chest x-rays12, we included similar data for males and females to assess potential sex-related interactions in the hypothesized airway changes associated with COVID-19.Accordingly, the primary objective of our study was to determine the relationship between central conducting airway diameter and COVID-19. This retrospective, case–control study used chest CT scans to test the hypothesis that people with COVID-19 would have larger central conducting airways than healthy controls. Additionally, because there are sex-related differences in airway size across the lifespanThis retrospective study was approved by the Institutional Review Board at the Mayo Clinic (IRB no. 17-008537) and conformed to the standards of the Declaration of Helsinki, except registration in a database. Images were collected as part of routine clinical care. Informed consent was waived as no identifiers were used, the data already existed, the research did not affect patient care and the patients’ parent/legal guardian did not opt out of their data being used for research. This consent waiver was approved by the Mayo Clinic Institutional Review Board.14, we did not include data from the Delta “wave” (B.1.617.2) and subsequent “waves” of SARS-CoV-2 variant predominance.Using three-dimensional reconstruction from low-dose high-resolution CT, we retrospectively assessed airway luminal cross-sectional area in patients with COVID-19 and in healthy, sex- and height-matched controls. The CT scans for people with COVID-19 were collected between March 2020 and August 2021. The healthy control cohort represents a historical reference group and CT scans for were collected before the COVID-19 pandemic between March 2009 and March 2018. Notably, because there were demographic and clinical differences in patients who tested positive for COVID-19 during different periods of SARS-CoV-2 variant predominance−2.The subject inclusion paradigm is displayed in Fig. n = 12 males, 8 females) were excluded due to poor quality images—defined by the assessor being unable to visualize all necessary airways. The final cohort of patients with COVID-19 consisted of 114 patients, including 48 males and 66 females.For the COVID-19 cohort, 1566 patients met the initial inclusion criteria, and their medical history was screened for the pre-specified exclusion criteria. After exclusions, 134 patients with COVID-19 met the criteria, and their images were analyzed for airway luminal area. During the analysis, 20 additional patients with COVID-19  who were included in our previous study were considered for inclusionght Fig. . The fin11, and are briefly described herein. A posterior-anterior and lateral topogram is obtained at 120 kV and 35 mA. Spiral acquisitions with a pitch of 1.2 are utilized. Kilovoltage is set at 120 with a standard milliampere-second value of 140. Post imaging reconstructions are obtained in the axial and coronal plane using a B46 kernel. Slice thicknesses of 1.5 mm and 3 mm are reconstructed. Maximal intensity projections in the axial and coronal planes are completed with a slice thickness of 10 mm and reconstruction increment of 2.5 mm. Our institution and this project used standardized CT algorithms. Images were acquired at end-inspiration after patients were requested to take a large inspiration and hold their breath. Notably, patients were not instructed to maximally inhale to total lung capacity, thus, lung volumes were not able to be matched between patients . The software algorithm isolates the airways from other tissue and creates a three-dimensional reconstruction. The cross-sectional area of the conducting airways was measured at three points  for each of the following airways: the trachea, right and left main bronchus, left and right upper lobes, intermediate bronchus, and left lower lobe. Anatomical bifurcations defined the proximal and distal point of the measured airways. Additionally, lengths of the trachea, right and left main bronchus, and intermediate bronchus were assessed.As previously describedDescriptive statistics are presented as mean ± standard deviation (SD) within the text and tables. Separate univariate analyses of variance were used to compare metrics of central airway size between patients previously diagnosed with COVID-19 and height- and sex-matched controls. Statistical models were performed in duplicate using two representations of luminal airway size— the measurement at the middle of the airway and the average of three measurements . Interpretation of findings did not differ between the statistical models, and findings using the average of three measurements of the airway are presented in the text. Findings using the measurement at the middle of the airway are presented in supporting information Tables  and S2.15 to predict luminal size of each airway. CHAID analysis builds a predictive model to determine the best cutoffs for the input variables to predict an outcome. CHAID creates all possible cross-tabulations for each categorical predictor until the best outcome is achieved and no further splitting can be performed.Given the known modifying effects of sex, analytical models were performed in duplicate with and without dichotomized models by sex. We also performed exploratory analyses on two subgroups of patients with COVID-19 dichotomized based on hospitalization status using univariate analyses of variance. Categorical variables  and patient characteristics  were used to construct a decision tree based on the exhaustive Chi-Square Automatic Interaction Detection (CHAID) algorithmp < 0.05. Analyses were performed using IBM Statistical Package for Social Sciences . Figures were created using GraphPad Prism software .Assumptions of normality were confirmed with Shapiro–Wilk tests and assumptions of homoscedasticity were confirmed with Levene’s test. Reported p-values are two-sided, and the interpretation of findings was based on p < 0.001).Patient characteristics are presented in Table n = 232). Sex and group (COVID-19 vs. control) were included in each decision tree, and four terminal nodes were employed—typified by a 2 × 2 contingency table. These data suggest that both sex and COVID-19 are predictors of airway luminal area.For all central conducting airways, luminal area was ~ 13% larger among males and females with COVID-19 compared to sex- and height-matched controls, Fig. p < 0.001; Tables 11, airway luminal area was ~ 30% greater in males than females . For both males and females, patients who were hospitalized with COVID-19 had greater lung volume and % predicted lung volume at which the images were obtained compared to patients who did not require COVID-19-related hospitalization. For the preponderance of airways, there were no differences in luminal size between patients COVID-19 stratified by hospitalization status, except for the left upper lobe for males and females and the right upper lobe for males.Although the primary objective was to determine the association between COVID-19 and luminal airway size, additional exploratory analyses were performed among COVID-19 patients based on hospitalization status. Twenty-two of 48 (46%) males with COVID-19 and 15 of 68 (22%) females with COVID-19 required COVID-19 related hospitalization Table , with thThe primary aim of this study was to evaluate central conducting airway size in people previously diagnosed with COVID-19 in comparison to healthy, sex- and height-matched adults. Consistent with our hypothesis, we found that both males and females previously diagnosed with COVID had larger luminal areas of central conducting airways compared to matched controls, including trachea, left and right main bronchus, intermediate bronchus, left and right upper lobe, and left lower lobe. Additional exploratory analyses also demonstrated that compared to patients who did not require COVID-19-related hospitalization, patients who were hospitalized with COVID-19 had no differences in luminal areas of central conducting airways. These findings suggest that symptomatic COVID-19 infection may be associated with pathologically larger central conducting airway luminal areas than healthy, sex- and height-matched controls.19. In the context of COVID-19, SARS-CoV-2 enters human bodies through respiratory droplets and attaches to epithelial cells of the conducting airways, suppressing the mucociliary apparatus that removes secretions and inhaled particles21. The resulting accumulation of contaminated mucous and airway edema may contribute to the increased luminal areas we observed on CT images of patients with COVID-1924. The disease course of COVID-19 in the conducting airways has yet to be elucidated. Previous studies demonstrate that airway luminal area may increase proportionate to disease severity and improve with resolution of acute infection25. Our study supports the finding of larger airways among patients with COVID-19 compared to controls, however, our study does not provide information on chronic changes to the airway following recovery from COVID-19. Airway luminal area is the major determinant of airway resistance and is particularly important when considering the implications of airway resistance in health and disease28. Although an increase in airway luminal area will decrease the resistance to airflow, this increase in airway luminal area will also contribute to reduced flow for a given pressure. Thus, the increase in airway luminal area can reduce the expectorant role of the conducting airways to clear mucous and debris before reaching the lung. Greater airway luminal area may also contribute to greater deposition of inhaled particles in lower portions of the airway29. Paradoxically, then, larger airway size may both predispose and be the result of inflammatory processes which ultimately result in mucous plugging and decreased airflow30.Disease processes can impart differences on the luminal area of the conducting airways, which transport gases without participating in gas exchange32. Males have larger luminal areas than females in the central conducting airways; this difference in airway size may affect resistance to air flow and aerosol deposition across the lifespan33. Consistent with previous studies, males in our cohort had larger luminal areas than females in all seven of the central conducting airways that were measured. However, there were no sex-related interactions in the differences observed in the luminal area of conducting airways among people with COVID-19 compared to controls. Thus, sex does not appear to interact with the observed effects of COVID-19 on central conducting airway luminal area.In addition to pathologic differences in the luminal area of conducting airways, anatomic differences between males and females are well established in the literature6 and CT reflect a typical lung injury of viral pneumonia8. The primary radiographic findings are ground-glass opacity and pulmonary consolidation—suggesting the possible presence of organizing pneumonia8. Although effective vaccines and therapeutics are available in many countries, about one-third of COVID-19 survivors have residual abnormalities on chest CT 1 year after COVID-1934. Thus, familiarity with sequelae of COVID-19 pneumonia on chest imaging may be important to evaluate potential causes of chronic residual abnormalities or respiratory symptoms after COVID-1936.Generally, radiographic findings associated with COVID-19 from both chest x-ray34. The larger airway luminal area may reflect traction bronchiectasis and could contribute to post-COVID respiratory symptoms37. Although no consensus currently exists for imaging management of patients with subacute COVID-19, our findings may suggest that enlarged central conducting airway luminal area may be a consideration in pulmonary sequelae among COVID-19 survivors.In this context, our findings showing larger central conducting airway luminal area among patients with COVID-19 may have clinical implications for post-COVID conditions—also referred to as “long COVID”, “long-haul COVID”, or “post-acute sequelae of COVID-19”38. Thus, although caution is required while interpreting absolute airway diameters, our primary comparison between cohorts is likely unaffected. Second, we used nonprobability sampling and a simplistic, cross-sectional design. Although our findings and those of others suggest a relationship between COVID-19 and larger airway luminal size39, the data should not be used to infer a definitive causal relationship or definitive temporal changes associated with COVID-19. Third, assessments of pulmonary function were not available in this cohort, as such, we were not able to determine the potential relationship between larger airway size and pulmonary function. Fourth, assessments of putative factors were not assessed, including, concentration of angiotensin converting enzyme receptors and changes in the pulmonary interstitium.Several limitations resulted from the design of this study, which may highlight areas for future investigation. First, the end-inspiratory lung volume was not standardized to total lung capacity. Rather, subjects were instructed to inspire and hold their breath. Notably, there were no observed differences in the relative lung volume between the two cohorts . Additionally, lung volume also has less of an influence on more proximal airways (which the current study assessed) compared with more distal airwaysOur findings suggest that males and females previously diagnosed with COVID-19 have larger luminal area of conducting airways compared to healthy sex- and height-matched controls. Further, COVID-19-related hospitalization was not associated with changes in luminal area of conducting airways among patients with COVID-19. A key limitation of the study is that the COVID-19 disease course was not characterized.Supplementary Information.


However, these products were designed initially for generic anticancer purposes and not specifically for BC treatment. With a better understanding of the molecular biology of BC, several novel and promising nanotherapeutic strategies and devices have been developed in recent years. In this context, multi-functionalized nanostructures are becoming potential carriers for enhanced chemotherapy in BC patients. To design these nanostructures, a wide range of materials, such as proteins, lipids, polymers, and hybrid materials, can be used and tailored for specific purposes against BC. Selective targeting of BC cells results in the activation of programmed cell death in BC cells and can be considered a promising strategy for managing triple-negative BC. Currently, conventional BC screening methods such as mammography, digital breast tomosynthesis (DBT), ultrasonography, and magnetic resonance imaging (MRI) are either costly or expose the user to hazardous radiation that could harm them. Therefore, there is a need for such analytical techniques for detecting BC that are highly selective and sensitive, have a very low detection limit, are durable, biocompatible, and reproducible. In detecting BC biomarkers, nanostructures are used alone or in conjunction with numerous molecules. This review intends to highlight the recent advances in nanomedicine in BC treatment and diagnosis, emphasizing the targeting of BC cells that overexpress receptors of epidermal growth factors. Researchers may gain insight from these strategies to design and develop more tailored nanomedicine for BC to achieve further improvements in cancer specificity, antitumorigenic effects, anti-metastasis effects, and drug resistance reversal effects.Breast cancer (BC) is a highly metastatic multifactorial disease with various histological and molecular subtypes. Due to recent advancements, the mortality rate in BC has improved over the past five decades. Detection and treatment of many cancers are now possible due to the application of nanomedicine in clinical practice. Nanomedicine products such as Doxil Breast cancer (BC) is one of the most prevalent cancers in women . This tyBC is classified into three main tumor subtypes rooted in enhancement in expression of either progesterone receptor (PR) or estrogen receptor (ER), and/or epidermal growth factor receptor 2 (ERBB2) gene mutation ,12,13,14The prognosis for women who have developed metastatic triple-negative BC is poor. The use of antibody–drug conjugates (ADCs) has been examined as a viable therapy option, especially in extensively pretreated illnesses. In metastatic triple-negative BC, sacituzumab govitecan is a new antibody–drug conjugate (ADC) that has demonstrated promising results. The FDA has designated it as a breakthrough therapy for treating individuals with previously treated metastatic triple-negative BC. For sacituzumab govitecan to operate, an antibody targeting the most common kind of BC-specific antigen, a human trophoblast cell surface antigen 2 (Trop-2), must be combined with the enzyme inhibitor SN-38 (topoisomerase I inhibitor) through a unique hydrolyzable linker ,20,21. SOne of the most important advances in treating metastatic BC has been the introduction of anti-HER2 drugs, which have considerably improved survival results. Notwithstanding their effectiveness, it is still necessary to use chemotherapy in conjunction with anti-HER2 monoclonal antibodies. When an antigen-specific antibody is coupled with a robust, cytotoxic payload, the outcome is an increased therapeutic index known as antibody–drug conjugates (ADCs). Creating antibody–drug conjugates, which consist of a cytotoxic agent and a monoclonal antibody carrier, provides a significant alternative to the traditional approaches used in chemotherapy. An anti-human epidermal growth factor receptor 2 (HER-2) antibody, a cleavable tetrapeptide-based linker, and a cytotoxic topoisomerase I inhibitor are the three components that make up the antibody–drug combination known as trastuzumab deruxtecan. Trastuzumab deruxtecan was found to have a promising efficacy in patients with HER2-negative or low-expressing disease, who have few treatment options due to the absence of the HER2 gene. This finding led to favorable outcomes in patients with HER2-positive BC who had undergone extensive prior treatment. The US Food and Drug Administration has awarded trastuzumab deruxtecan rapid approval in advanced or unresectable HER2-positive BC that has been treated with at least two HER2-targeting therapy lines, based on results from recent clinical studies. It is feasible that newer HER2-targeted ADCs might offer a wide range of novel therapeutic uses outside typical HER2-positive BC because of their enhanced pharmacological features, such as the ability to target cells with low expression of the HER2/ERBB2 mutation ,25. FurtThere are two options for the surgical phase with similar survival rates: a lumpectomy with radiation if the tumor can be excised completely with good cosmetic results, or a mastectomy ,10. TherFor health care professionals, screening and diagnosis methods are crucial to supply individualized therapies that will improve outcomes and survival ,41,42. CAs conventional cancer diagnostics and therapeutic tools become less effective due to increased systemic toxicity, nanotechnology is currently utilized to improve diagnosis and mitigate disease severity ,53,54. ANanotechnology-based immunotherapy agents have been employed for multiple cancer types to shrink the malignant cancerous cells and preserve the benign cells at the target spot ,65,66,672Te NPs and their utilization in the biomedical situation demanding the maximum NP dosage . This study’s evidence highlighted the impending utilization of Ag2Te NPs in the biomedical arena and as X-ray contrast agents for BC detection [Multipurpose nanotechnology-based plans with diagnostic imaging and targeted treatment roles have been among the most debated topics in the nanomedical research field ,71,72,73etection .Limitations in conventional diagnostic approaches have resulted in the emergence of novel means, such as nanotechnology, which is expected to positively impact BC patients’ survival and quality of life. There is a level of dependency on estrogen for growth in around 70–75% of BCs, which is shown by the expression of the estrogen receptor (ER) . In factAs discussed earlier, BC is the most widely encountered malignancy with a high mortality rate . The usuRecently, biosensors have been widely explored in the recent era for diagnosing malignancies . BiosensVarious biomarkers have been exploited during the initial diagnosis of BC . The metHER/neu, a human epidermal growth factor receptor 2, has been recognized as a critical determinant in malignant alterations in BC that can indicate its destitute prognosis . SpecifiThere has been extensive research on using nanotechnology for early cancer detection ,99.Due to its unique surface characteristics and simple surface functionalization, many applications of graphene and its derivatives have been introduced into the biomedical field . Graphen2O2 in clinical BC analysis. The nanocarrier system exhibited high sensitivity and good biocompatibility for instantaneous tracing of H2O2 released from varying BC cells [Xu and coworkers have investigated nano-enzyme decorated graphene quantum dots for electrochemical recognition of HBC cells . GraphenIn addition to chemotherapy, photodynamic therapy (PDT) and photothermal therapy (PTT) can be used as a non-invasive therapeutic strategy as a complement to overcome the deficiency of monotherapy. As a result of light exposure, photoactive therapeutics (photosensitizers) can produce reactive oxygen species (ROS) or hyperthermia, thus killing cancer cells. Mesoporous silica NPs are suitable for multi-drug loading since they have a large surface area, pore size, and volume, making them a suitable target for photo-chemotherapy. The structural merits of MSNs have generated considerable attention as potential partners for PDT in recent years. In solid tumors, many photosensitizers aggregate easily, reducing their efficacy, and have poor intracellular uptake, making them unsuitable for use. The incorporation of MSNs prevents PSs from aggregating and improves their targeting ability and biocompatibility, resulting in fewer side effects and stronger anti-cancer effects.Wang and coworkers have proposed mucin-1 protein (MUC-1) targeted magnetic silica-based mesoporous NPs to capture MFC-7 cells. These silica NPs were coupled with a folate-receptor-directed fluorescent probe by conjugating folic acid and fluorescein isothiocyanate over the surface of bovine serum albumin for selective and specific labeling of folate receptors in HER-2over-expressed MCF-7 cells. The quantitative assay developed showed superior specificity and sensitivity towards MCF-7 cells . Chen anQiao and coworkers have explored pH-responsive plumbagin-loaded theranostic NPs composed of mesoporous silica-covered gadolinium-III, conjugated with zoledronic acid for early recognition of BC-associated bone metastasis with increased sensitivity and selectivity .Salahandish and coworkers have demonstrated gold and silver NPs grafted graphene and nanostructured polyaniline for highly selective biosensing and a wide linear response range. The nanostructures were found to be ultrasensitive for label-free sensing of BC cells. These nano-biosensors had a faster response rate and a detection limit as low as 2-cells/mL for the SKBR-3 cell line. The sensor also exhibited a high detection efficiency of 90% .In another experiment by Saeed et al., developed an electrochemical DNA biosensor based on AuNPs-modified graphene oxide for simultaneous detection detect CD24, a new prognostic marker in BC, and a transmembrane protein tyrosine kinase (ERBB2). The developed DNA nanosensor utilized a sandwich-type detection method, and amperometric detection was used to assess the sensor’s response. For this purpose, the gold nanoparticles were grafted onto the glassy carbon after being attached to graphene oxide. Surface immobilization was used to attach thiolated nucleic acid capture probes. Because of the amplification of the signal that was obtained, sensitive detection of both BC markers (ERBB2c and CD24c) was possible. The detection limit was found to be 0.16 nM and 0.23 nM for ERBB2 and CD24, respectively . We beli3N4). Cu2ZnSnS4 NPs can be used for the detection of HER-2 in BC [Dong and coworkers have investigated HER-2 functionalized magnetic gold-shelled poly (lactic-co-glycolic acid) nanocarriers for ultrasound or magnetic resonance imaging and BC photothermal therapy. The nanocarrier system exhibited a receptor-specific binding HER-2 overexpressed in BC cells (SKBR-3) with a significantly high binding affinity. In vitro studies revealed that NPs possessed both enhanced ultrasound and magnetic resonance imaging properties . Phototh-2 in BC .Tian and associates have investigated AuNPs for electrochemical sensing (label-free) of microRNA-21 (miRNA-21) via a redox indicator. These sensors exhibited high sensitivity, selectivity, and reproducibility in blood serum samples . Feng anArteaga and coworkers have utilized silver NPs (citrate reduced) as a substrate for surface-enhanced Raman spectroscopy to quantify BC-associated elevation in sialic acid in saliva. This simplified test exhibited a 94% sensitivity and 98% specificity for the diagnosis of the BC patient, with a cut-off concentration of sialic acid assumed to be >7 mg/dL .In another study, label-free geno-sensors were developed for the detection of BC-specific biomarker miRNA-21. The sensor was based on graphene functionalized silver NPs, resulting in significant electrochemical signal amplification. This biosensor was found to be very reproducible in assessing blood samples and can be used for direct recognition of miRNA-21 during the early stages of BC without the need for any sample preparation or RNA extraction .Albernaz and coworkers have investigated nano-radio-labeled superparamagnetic iron oxide NPs. These NPs were further conjugated with diethylene triamine penta-acetic acid labeled with technetium 99 and gallium 68 for SPECT and PET. The results demonstrated a high accumulation of nanocarrier systems in tumorous cells compared to normal cells, giving a clear contrast image by both techniques (SPECT and PET) .Hsu and coworkers have proposed a multimodal imaging probe, called “all in one NP (AION)”, which is developed by combining near-infra-red fluorophore, silver sulfide NPs, and iron oxide NPs into PEGylated micelles. The AION exhibited a minimal release of silver ions leading to a significant decrease in associate cytotoxicity. A strong contrast was generated compared to all imaging modalities and exhibited a strong contrast when injected intravenously for in vivo tumor imaging. Thus, AION was suggested as an excellent candidate for BC detection with various imaging opportunities .In 2021, Li and colleagues developed a motif (RXDLXXL)-linked arginine-glycine-aspartate nano-peptide and conjugated it with superparamagnetic iron oxide NPs (cFK-9-USPIO) for molecular imaging of integrin protein (αvβ6), overexpressed in BC. In comparison to the controls, the in vivo MRI of four nude mice carrying T1 xenograft tumors revealed a significant decline in T2 signal intensity in the BC tissue. Prussian blue staining provided additional confirmation that v6 integrin-targeted NPs had specifically accumulated in 4T1 BC cells. Comparatively, significantly fewer particles were seen in the 4 T1 tumors of mice that had been injected with control USPIO . We beliIn another study, ultra-small iron-oxide NPs coupled with BRBP-1 were investigated for NIRF and MRI of BC-associated brain metastasis. The peptide-modified nanocarrier system was found to enhance the imaging signal induced by targeting the ability of BRBP-1, thereby increasing the potential of nanocarriers for the diagnosis of brain metastasis associated with BC .Semkina and coworkers have studied anti-vascular endothelial growth factor coupled iron-oxide NPs intended for targeted transport of DOX in the murine breast adenocarcinoma 4T1 cell line. After 24 h, MRI was used to study how NPs accumulated in cancer cells, demonstrating that the carrier system can simultaneously deliver drugs and diagnose cancer . We beliPacheco and coworkers have proposed molecularly imprinted polymeric sensors to sense and quantify HER-ECD. The quantification and detection limits were 1.6 ng/L and 5.2 ng/mL, respectively. The biomarker was found to be more selective in comparison to other protein biomarkers . Zhang aJin and coworkers have developed a peptide (cyclic arginine-glycine-aspartic acid) conjugated polymeric NPs using poly(2-methoxy-5-(2-ethyl-hexyloxy)-1,4-phenylene vinylene) as a photosensitizer targeting TNBCs. The polymeric nanoconjugates exhibited bright flourescence, increased stability, and could generate reactive oxygen species on light irradiation. It was demonstrated that such nanoparticulate systems are useful as diagnostic tools for clinical use .Zhang and coworkers have explored tumor-derived exosomes as biomarkers for detecting MCF-7 cells by targeting overexpressed mucin-1. The proposed on–off apta-sensors were turned on in the presence of mucin-1 on tumor-derived, resulting in fluorescence signal emission. The exosome-based biosensor successfully quantified a small number of samples, making them highly sensitive sensors . Xu and 2O2 liberated from BC cells. The bismuth selenide NPs were regarded as electrochemical signal boosters, while the Au coating added to the stability of these signals. The proposed biosensors offered a low detection limit (10 × 10−9 M) along with the excellent capacity to distinguish two variations of BC cells (MFC-7 and MDA-MB-231) contingent on the variation in H2O2 generation [Mohammadniaei and colleagues have developed a new electrochemical biosensor using a topological illustrator and metallic DNA. Bismuth selenide NPs were prepared and sandwiched between an Au electrode, and Au deposited a thin layer of bismuth selenide. This was followed by immobilizing eight silver ion-mediated dsDNA onto the substrate to detect Hneration .Multifunctional nanocarriers include polymeric NPs, self-nanoemulsifying drug delivery systems (SNEDDS), liposomes, and mesoporous inorganic NPs. The nanocarriers are supposed to permeate through the intestinal mucosa ,135,136.The multi-functional nanocarriers can release the drug in a specific redox-triggered environment for intra-tumoral drug release, as shown in Cancers of the breast are the world’s deadliest diseases, with a higher death rate among women. BC treatment is often associated with using oral chemotherapeutics in terms of radiation therapy, chemotherapy, hormonal therapy, and multi-functionalized ligand-mediated therapy . AdditioFor the development of NPs, Zhang and associates  used eleL. Palanikumar and the research group developed biodegradable pH-responsive NPs conjugated with a polymeric system comprising poly(lactic-co-glycolic acid (PLGA). This polymeric system was then coated with bovine serum albumin (BSA). The important feature of this formulation was the inhibition of the macrophages, which causes the inhibition of recognition targets. Moreover, after the uptake of NPs, intracellular microenvironment conditions lead to degradation of NPs and proficient anti-cancer activity against BC cell lines .ε-poly-l-lysine-DOX/lapatinib polymeric nanoplatform for encapsulation of anti-cancer drug lapatinib for its conversion into switchable charge based dual pH-responsive NPs. Advanced physicochemical properties of novel NP conjugates lead to stability in the circulation of physiological conditions. However, charge switching capability from negative to positive charge leads to high-level sensitivity in the slightly acidic TME, facilitating strong muco-penetration [Similarly, Zhihao Guo and co-workers developed a tailor-made 2,3-dimethyl maleic-anhydride-poly(ethylene glycol)-etration .Liu and coworkers  developeMoreover, the ionization of PHIS in the TME converts its hydrophobicity into hydrophilicity, which is very helpful in exerting immune regulation. Hydrazone bond breakage at endosomal pH accelerated the release of DOX, exerting cytotoxic effects. HA-DOX was overexpressed in BC cells and resulted in the internalization and inhibition of cell growth. In summary, this multi-functionalized nano-core system could deliver precisely in the TME and BC cells to achieve synergistic effects for excellent tumor-targeting ability against BC .Carbon-based NPs can be synthesized via different approaches based on their application . TherefoAdditionally, Abid Hussain and Shengrong Guo investigated a new controlled system to release drugs from mesoporous carbon NPs (MCNs). The surface of the NPs was decorated with natural sophorolipids (SLPD). The pores were developed on the MSNs for inducing photothermal activity and encapsulating a chemotherapeutic drug (DOX). This advanced nanoparticulate system acts as a checkpoint for trapping DOX inside the pores of mesoporous carbon NPs and triggering its release in the TME via NIR irradiation. As a result, they demonstrated that this system is highly effective against BC cell lines .In 2019, Batool and collaborators synthesized SNEDDS of tamoxifen (Tmx) for targeting BC . FollowiMoreover, we believe that solid SNEDDS can improve patient compliance and tackle the limitations associated with liquid SNEDDS capsules. However, the selection of SNEDDS components is heavily influenced by their physicochemical features, drug solubility, and physiological fate.3BO3) at a molar ratio of 1:6 to convert them into a powdered form. These precursors were then heated under airflow in the horizontal furnace. The resulting crude samples were then centrifuged to remove impurities, followed by scattering in the presence of 25 mL of H2O2 and then transferred to a Teflon-lined autoclave for thermal treatment at 120 °C for 24 h. Furthermore, the dispersions were cooled at room temperature and subsequently centrifuged at 4000 rpm for 10 min, followed by vacuum drying overnight, resulting in the synthesis of BNNPs. These NPs have been characterized via vitamin C triggered degradation, cellular uptake, PET imaging, biodistribution in tumor-bearing mice, degradation behavior in vitro, degradation behavior in vivo, tumor-bearing animal models, radiolabeling, in vivo Boron Neutron Capture Therapy (BNCT), and histological studies. Results concluded that in PET imaging, the coated BNNPs exhibited high tumor boron accumulation while maintaining an excellent tumor to non-tumor ratio, and BNNPs were found to be rapidly cleared from major organs according to ex vivo ICP-OES analysis. Furthermore, when compared with the control group, animals treated with BNNPs showed tumor growth suppression with negligible side effects. This novel strategy not only employed the high boron content of BNNPs but also efficaciously completed an on-demand degradation of BNNPs to avoid the potential toxicity caused by the long-term accumulation of NPs [In 2019, Le and coworkers developed the advanced and on-demand technique of biodegradable boron nitride NPs (BNNPs) for specified targeting against negative BC cells via neutron capture therapy of boron . For then of NPs .1H-NMR and 31P-NMR spectroscopy, DLS, zeta potential, TEM, and atomic emission spectroscopy. Based on the findings of this research, synthesized nanocarriers are potentially useful in future treatments for BC [Overexpression of estrogen hormone is linked to BC, and anti-estrogen therapies involving tamoxifen are the most commonly prescribed methods of treatment for it ,159,160.s for BC . Yi Zhans for BC .The functionalization of liposomes with monoclonal antibodies is a potential strategy for increasing the targeting capability of the HER2, overexpressed in HER2-positive BC cells ,165,166.We already know that anticancer medications can be delivered to the body via liposomes to either lessen the harmful effects of the drugs when administered on their own or lengthen the circulation time and effectiveness of the treatments . AccordiVarious preclinical studies regarding NPs against BC involve methotrexate and beta carotene conjugated lipid polymer hybrid NPs for inducing BC by A Jain and co-workers . MoreoveMore accurate and targeted co-delivery of both therapeutic and diagnostic compounds is being pursued due to the last two decades’ worth of nanotechnological breakthroughs in biomedical research for cancer treatment using drug delivery carriers and contrast agents . Nano-caA class of nanomaterials that are inert (resistant to corrosion/oxidation) and have distinct physical and sensing characteristics are inorganic NPs. For the identification of a variety of biomarkers, inorganic NPs, in particular silver and gold nanoparticles (AuNPs and AgNPs), are very precise and sensitive biosensors . For exaProteins, nucleic acids, and other macromolecules are dephosphorylated by the enzyme alkaline phosphatase (ALP). It has the potential to be a biomarker for many different illnesses, including hepatobiliary, osteopenia, and breast cancer. In light of this, a colorimetric sensor for the ALP test was developed. This sensor is based on the enzyme’s ability to dephosphorylate the compound p-aminophenol phosphate (pAPP) into pAP. Silver nanoparticles (AgNPs) and Ag+ ions are present in a solution that has been made using a low concentration of NaBH4, and pAP mediates the growth of AgNPs by lowering the concentration of Ag+ ions to increase the intensity of localized surface plasmon resonance because pAPP is unable to cause a reduction of the remaining Ag+ because the hydroxyl is being covered by phosphate. The colorimetric detection of the pAP-mediated development of AgNPs in the presence of an ALP served as a proof of concept for the quantitative analysis of the ALP. AgNPs’ extremely sensitive enzymatic growth offered a lower limit of detection of 0.24 U/L and a larger dynamic linear range of 0.5–225 U/L than had previously been reported. For the ALP test in human blood, the application of pAP improved the sensor’s selectivity, resulting in a high recovery rate and high accuracy of 99.2 1.5 percent for the traditional addition technique .For instance, a liposomal layer was recently added to AL (LAL), followed by radiolabeling and the addition of polyethylene glycol (PEG), to create a theranostic dual-layered nanocomposite . LAL’s iIn a related investigation, the effectiveness of hybrid liposomes (HL) as therapeutic agents and the capability of Indocyanine Green (ICG), a fluorescent probe containing HL, to identify malignancy in orthotopic graft model mice of BC (MDA-MB-453) were assessed. As a result of their therapeutic benefits and their capacity to identify (diagnose) cancer in an orthotopic graft model mice of BC, the results suggested that HL and HL/ICG might be theranostic targets .Due to their excellent characteristics, conjugated polymer NPs (CPNs) have become a novel and promising group of BC theranostic weapons . In thisA multimodal polymeric contrast agent and imaging techniques have the potential to make important advances in the area of biomedicine . To addrWe believe that patients diagnosed with BC benefit from an improved prognostic evaluation when multimodal imaging and therapy features are combined. The above dual systems worked fine and can be applied for BC imaging and treatment.Advancements in nanomedicine have brought nanotechnology-based methods to the forefront of cancer research. Multi-functionalized nanostructures with intelligent design and unique physicochemical properties have revolutionized targeted drug delivery and shown great potential in BC theranostics. An overview of BC nanomedicine’s experimental and clinical accomplishments using active targeting and flexible biomedical nanostructures is presented in the paper. To give an example  and their potential applications in BC imaging, drug delivery, combination therapy, and theranostics. Furthermore, it can be concluded that nanovesicles such as liposomes, nanogels, polymeric nanocarriers, mesoporous inorganics, biodegradable boron nitride, self-nanoemulsions, core–shell NPs, etc. have set a benchmark in the prevention of multiple BC subtypes by targeting drug delivery. Even though preclinical results indicate that these nanostructures are clinically relevant, several problems still exist and need to be tackled before their clinical application can be sought. ®) exhibits a prolonged half-life, increased tumor drug concentration, and superior antitumor efficacy compared to conventional DOX [Moreover, the surface properties of a nanosystem determine its behavior and interaction with proteins and cells . Severalonal DOX . TargetiBC mortality rates can be significantly reduced by early detection of the disease with nanotechnology and therapeutics that are more effective. Nanomedicine promises several opportunities for both specific and multi-acting cancer treatments; nonetheless, the most important question is whether or not nanomedicine will be clinically transferrable in the near future. Clinical practice for BC has not been enhanced by nanostructures as expected. The main reason may be the absence of adequate preclinical models that effectively mimic, both spatially and physiologically, actual BC and its intricate interactions with the surrounding microenvironment. Because the vast majority of potentially effective drugs and therapeutic approaches developed in preclinical studies do not reach clinical trials, clinical translation cannot be carried out separately from this point. For clinical approval to happen more rapidly, a collaboration between academia and pharmaceutical companies will be beneficial to developing the field and improving clinical trials. Examining questions of long-term toxicity, biocompatibility, and atomic-level organization of these nanostructures is equally crucial before entering the market. Hopefully, cancer nanomedicine will enter its next phase of advancement by incorporating nanostructures into rationally designed therapies. As a result, new opportunities in BC nanomedicine may arise by developing innovative, high-level strategies and systems based on active targeting using a variety of nanostructures.In our opinion, BC is mostly hormone-sensitive, employing both estrogen and progesterone sensitivity. Therefore, targeting the receptors of specific hormones through the coating of targeted ligands is the most effective method of halting BC tumor growth as an inducer for binding specifically to the receptor site. Additionally, shifting from traditional chemotherapy to a more effective targeted endocrine therapy can have a substantial impact. In this regard, we can safely assume that synergistic conjugation of chemotherapy and endocrine treatment will impart a significant effect and prove BC treatment as a breakthrough. Using nanoscale compositions of materials that have already been approved for use in humans is expected to be the first step in developing potential future nanomaterials in BC treatment. Before the significant potential of nanomedicine can be realized for the benefit of the affected patients, ongoing interdisciplinary collaborations between cancer biologists, toxicologists, bioengineers, material scientists, medical oncologists, and pharmacologists will be required.


Background: Extended-spectrum β-lactamase (ESBL) producing organisms are resistant to a wide range of broad-spectrum antibiotics, and their emergence is a significant driving force of antibiotic resistance. Most South-Asian countries have become hotspots for antibiotic resistance, so specifics of ESBL data are critical to tackling antibiotic resistance. We present the temporal changes in ESBL-producing organisms cultured in our tertiary care referral centre.Methods: Over a year, a historical cohort analysis was carried out at our tertiary care referral centre in Southeast Asia. Samples from patients presenting with acute surgical conditions were sent for culture and sensitivity. The phenotype of all specimens was checked using the combination disc method. Antimicrobial susceptibilities to various antibiotics were also checked as per CLSI  guidelines.Escherichia coli (70%), Klebsiellapneumoniae (18%), and Pseudomonas aeruginosa (16%). The percentage of ESBL-producing organisms was 54.7%, which is significantly higher than in previous reports. Widespread resistance was found against commonly used antibiotics, including co-amoxiclav (81.9%), ceftriaxone (75%), ciprofloxacin (47%), and levofloxacin (35.7%). Sensitivities to combination antibiotics like piperacillin-tazobactam (79.2% sensitive), cefoperazone-sulbactam (84.3% sensitive), and imipenem-cilastatin (91.1% sensitive) were also noted to be falling.Results: Specimens from 170 patients were analysed. The mean age was 44.73±19.89 years, and there was a female predominance of 62%. The most common organisms were Conclusion: The incidence of ESBL-producing organisms continues to increase at an alarming rate, which mandates strict antibiotic stewardship and amendments to local guidelines. Antimicrobial resistance is one of the forefronts of battle in current medical practice. Increasing resistance to common as well as uncommon antimicrobial agents is alarming, and this problem has been declared a global threat to public health . South AEscherichia coli, Klebsiella pneumoniae, and Pseudomonas aeruginosa, were traditionally treated with penicillins, cephalosporins, and fluoroquinolones. After the acquisition of β-lactamases and extended-spectrum β-lactamases (ESBL) by these microbes, clinicians have been struggling to treat them even with broad-spectrum antibiotics like third-generation cephalosporins. This is a cause for concern as the choice of antibiotics to treat these multi-drug-resistant organisms is becoming very limited, and there might not be any viable antibiotics left if the problem is not addressed emergently [Gram-negative bacteria (GNB), particularly ergently .Whereas β-lactamases were only resistant to Penicillins and first- and second-generation cephalosporins, mutations in β-lactamases resulted in the evolution of ESBL, which provides resistance to third-generation cephalosporins as well as monobactams . The detA historical cohort analysis was carried out. Ethical approval was obtained from the local ethics committee . Specimens sent for culture and sensitivity underwent Gram staining after initial inoculation in blood and MacConkey agar for wound and pus specimens, and cystine lactose electrolyte-deficient (CLED) agar for urine specimens. All Gram-negative organisms had their antibiotic susceptibilities checked by the Kirby-Bauer disc diffusion method on nutrient agar with standard antimicrobial discs as per Clinical and Laboratory Standard Institute (CLSI) guidelines. A total of 170 cultures were included from January 2019 to June 2020. All cultured organisms were checked for antimicrobial susceptibility against Penicillins, first-, second-, and third-generation cephalosporins, fluoroquinolones, carbapenems, aminoglycosides, and tetracyclines. Representative drugs were used from the above classes in all cultures. ESBL production was confirmed by using the combination disc method . Organisms were phenotypically considered ESBL-producing when the zone of inhibition was ≥5 mm larger around the combination discs as compared to antibiotic discs. Data analysis was performed using IBM SPSS version 22. A chi-square test was used to compare frequencies, and an unpaired student t-test was used to compare means.Specimens from 170 patients were included in the study. There was a female predominance 62%). The mean age was 44.73±19.89 years. The distribution of microbes isolated from these cultures is elaborated in Figure %. The meOur study showed high levels of resistance to common antibiotics like co-amoxiclav (81.9% resistant), ciprofloxacin (47% resistant), levofloxacin (35.7% resistant), and ceftriaxone (75% resistant). Isolates were found to be sensitive to combination antibiotics like piperacillin-tazobactam (79.2% sensitive), cefoperazone-sulbactam (84.3% sensitive), and imipenem-cilastatin (91.1% sensitive), while aminoglycosides like amikacin and gentamicin also showed a reasonable sensitivity rate of 85.9% and 64%, respectively.Another interesting finding in our study was that the number of resistant organisms to co-amoxiclav, ciprofloxacin, levofloxacin, tazocin, and gentamicin was significantly higher for ESBL-producing organisms as compared to non-ESBL-producing organisms. This is illustrated in Table ESBL production by Enterobacteriaceae renders them resistant to even third-generation cephalosporins. Over time, there has been a rise in the number of ESBL-producing Enterobacteriaceae because of antibiotic resistance, as our study demonstrated that more than half of the isolates in our study were ESBL-producing. Our results show a significant rise in the multidrug resistance of ESBL-producing GNBs, with particularly high resistance rates to commonly used antibiotics including co-amoxiclav (89.4%), ciprofloxacin (60.8%), levofloxacin (44.4%), gentamicin (37.8%), and rising resistance rates for broad-spectrum antibiotics like piperacillin-tazobactam (17%). The proportion of ESBL-producing GNBs has also risen at an alarming rate to 54.7%.Antimicrobial drug resistance is an alarming concern in current medical practice. This alarming issue, especially in regions with poor antimicrobial stewardship such as South Asia, limits the efficacy of stronger antibiotics. The use of antimicrobials is one of the contributing factors to the development of antimicrobial resistance, and some authors have suggested that over-prescription of antimicrobials may be responsible for the increasing levels of antimicrobial resistance over time .Comparing our results with previous studies from the same region shows that the frequency of ESBL-producing Enterobacteriaceae has increased significantly. Jamil et al. cultured 75 isolates from 240 urine samples and reported an ESBL-production frequency of 25 (33.3%) . Khan etA comparison with international reports from different parts of the world shows varying numbers regarding the prevalence of ESBL producers in different geographical regions. Flokas et al. in their meta-analysis of pediatric urinary tract infections reported a pooled prevalence of 14% (95% CI: 8-21) among included studies, with a 12% (95% CI: 2-31) estimated prevalence from Europe from five studies and an estimated prevalence of 5% (95% CI: 0-7) in the Americas from three studies . There iThe susceptibilities of ESBL to various antibiotics showed an alarming level of resistance to commonly used drugs, while higher-level antibiotics such as carbapenems showed adequate levels of sensitivity when used in combination. The levels of resistance to other drugs are also on the rise, with 30% of the isolates being resistant to gentamicin. Other drugs like Linezolid have been used injudiciously in recent times, which has resulted in an alarming rate of resistance against this antibiotic; our data showed that 63% of the tested isolates were resistant to Linezolid. This is an alarming finding as Pakistan has already failed to achieve the targets set by the WHO’s 2017 mission report on antimicrobial resistance as per the global action plan of the World Health Assembly 2015.ESBL-producing organisms can be tackled by judicious use of broad-spectrum antibiotics, increasing the rate of cultures, and sparing antibiotics that have high levels of sensitivity. Carbapenem-sparing strategies are being practiced in some parts of the world to preserve some antibiotics as a last line of defence against ESBL-producing organisms . EmergenOur study showed a significant increase in the frequency of ESBL-producing organisms as compared to previous studies. In our study, more than half of the cultured organisms were found to be ESBL-producing with high levels of resistance to commonly used antibiotics. Emergent steps are needed to contain the rapidly rising resistance levels among bacteria, particularly in South Asia. These steps should include strict antimicrobial stewardship, judicious prescription of broad-spectrum antibiotics, and physician training to improve the practice of culturing organisms and the prescription of targeted antibiotics after cultures. These steps must be undertaken immediately to preserve the efficacy of antibiotics and tackle the menace of antibiotic resistance.


Background: Wastewater-based epidemiology (WBE) has the potential to inform activities to contain infectious disease outbreaks in both the public and private sectors. Although WBE for SARS-CoV-2 has shown promise over short time intervals, no other groups have evaluated how a public-private partnership could influence disease spread through public health action over time. The aim of this study was to characterize and assess the application of WBE to inform public health response and contain COVID-19 infections in a food processing facility. Methods: Over the period November 2020–March 2022, wastewater in an Arizona food processing facility was monitored for the presence of SARS-CoV-2 using Real-Time Quantitative PCR. Upon positive detection, partners discussed public health intervention strategies, including infection control reinforcement, antigen testing, and vaccination. Results: SARS-CoV-2 RNA was detected on 18 of 205 days in which wastewater was sampled and analyzed (8.8%): seven during Wild-type predominance and 11 during Omicron-variant predominance. All detections triggered the reinforcement of infection control guidelines. In five of the 18 events, active antigen testing identified asymptomatic workers. Conclusions: These steps heightened awareness to refine infection control protocols and averted possible transmission events during periods where detection occurred. This public-private partnership has potentially decreased human illness and economic loss during the COVID-19 pandemic. The COVID-19 pandemic has rattled life’s normalcy, from personal and institutional health to the food and commodity supply chains , in largArizona is one of the largest agricultural producers in the United States and contains over 26 million acres of farmland . The corTo prevent human and economic impacts from SARS-CoV-2 outbreaks in the agriculture sector in Arizona, the University of Arizona Yuma Center of Excellence for Desert Agriculture (YCEDA)—in coordination with Yuma County Public Health Services District (YCPHSD), the Regional Center for Border Health, Inc., and the Arizona Department of Health Services (ADHS)—launched a wastewater-based surveillance (WBE) system to monitor SARS-CoV-2 RNA for public health response across public and private sectors, including agriculture. This partnership was formed to maintain the wellbeing of the worker population, primarily in addition to the food processing facility. Although the study is not the first to report building-level sampling or performing WBE for public health benefit that either promotes subsequent clinical interventions and educational practices for infection control ,14,15, tBetween November 2020 and March 2022, one food processor facility’s wastewater was monitored for SARS-CoV-2 RNA. The facility is located in Arizona, approximately 30 miles from the United States-Mexico border, a company with annual revenue of over $30 million and customers in over 30 countries worldwide. During the study period, the monitored facility employed 277 workers, of which 246 were food handlers, 27 were administrative, and four were human resources professionals. Most workers were White, Hispanic (99%), and male (57%), with a mean age of 44 years old. Employees worked 8-h shifts, donned gloves when handling food products, and were required to follow public health guidelines  on-site.n = 41) with intact human coronavirus 229E. The mean recovery efficiency rates ranged between 6.81% to 31.60%. A standard curve (100–107 GC/L) for the N1 assay characterized RT-qPCR performance, including the slope (−3.39), y-intercept (41.37), and efficiency (97.4%). The limit of detection (2.00 × 103) and the limit of quantification (3.60 × 103) were reported per liter. The methodology is further detailed in a previous manuscript published by this group [Wastewater was collected at the in-flow of a concrete septic tank , fed solely by the facility’s plumbing. At least once every three-to-four days, a 1-L sample of raw sewage was collected from the septic tank. Samples represented 10-h composites since the septic tank pumped and flushed after reaching capacity, approximately every 10 business hours. The septic tank was also flushed after each event where individuals were diagnosed with SARS-CoV-2 on-site. SARS-CoV-2 particles were vacuum filtrated and concentrated via Centricon Plus-70 centrifugal filters, nucleic acids were extracted, and the N1 gene was enumerated via reverse transcriptase and quantitative PCR (RT-qPCR). Each collected sample was analyzed in triplicate, and the determination of a positive wastewater sample was based on the mean PCR quantification cycle. We spiked a subset of community wastewater samples (is group .q) < 37, while Cq values from 37–40 were considered indeterminate [q < 40; this resulted in several samples being interpreted as ‘positive’ despite being below the estimated limit of detection (LoD). All results—both positive and negative—were communicated with the food processing facility.Although the ideal interpretation would estimate absolute case numbers via calculations using the concentration of RNA in a given volume of wastewater, the individual waste-shedding rate still has not been fully elucidated ; therefoerminate . AlternaUpon finalized reports, positive samples triggered an immediate public health response that consisted of auditing and reinforcement of infection control protocols, including hand hygiene, masking, social distancing, and physical barrier implementation in communal environments . Similarly, facility leadership heightened awareness of disease prevalence among employees during morning and afternoon huddles to reinforce infection control protocols until wastewater samples were negative via RT-PCR. While follow-up clinical testing and vaccination services could not always be provided due to limited resources, in several cases, the facility implemented a point prevalence survey for SARS-CoV-2 infection via rapid antigen testing in addition to offering the Moderna SARS-CoV-2 vaccine for eligible employees. Three COVID-19 antigen tests were deployed based on accessibility: Abbott BinaxNOW, BD Veritor, and Care Start. However, individuals who administered tests did not document in what quantities each test was used. During these five events, all agricultural workers were tested via nares swab samples collected within 24 h of the positive wastewater result. Positive individuals and their close contacts were furloughed until deemed safe to return to work, in compliance with the contemporary United States Centers for Disease Control and Prevention (CDC) guidance. After the identification and isolation of positive individuals through clinical testing, wastewater samples were collected and tested within 24–72 h to determine if any individuals had residual viral shedding. If wastewater tested positive for SARS-CoV-2 RNA in subsequent collections, employees and wastewater were tested again, and so on; the entire surveillance and response structure is illustrated in From November 2020 onward, wastewater samples were collected consistently on a biweekly basis, except during the facility’s off-season, between 1 July 2021 and 2 August 2021, when the food processor facility suspends operations annually. During the study period, the Wild-type and variants of the SARS-CoV-2 virus circulated in the surrounding region. Therefore, we have contextualized wastewater and clinical testing results within the larger scope of the predominant SARS-CoV-2 variants experienced by the United States population, using two CDC MMWR manuscripts ,19,20; tWastewater was positive for SARS-CoV-2 RNA on 18 distinct sampling days  from a tq = 35.66; N1 = 3.9 log10 GC/L). On the following day, all 191 individuals who worked on-site in the previous week underwent rapid antigen testing, among whom four individuals tested positive  and were all asymptomatic. The four positive workers and one positive worker’s spouse—also employed by the food processing facility and tested negative—were furloughed the same day. The wastewater basin was flushed later that day. On the next business day, 7 December, the wastewater was tested again, and no SARS-CoV-2 RNA was detected. No workers reported symptoms during the following days after the removal of the first four individuals from the population. The negative wastewater samples suggested that all infected employees had been identified and removed from working in the facility.The morning of Wednesday, 2 December 2020, marked the first positive wastewater sample . All workers who were staffed on-site that week (n = 186) participated in antigen testing the same day. One agricultural worker tested positive for SARS-CoV-2 and was furloughed; this individual was asymptomatic. The wastewater basin was flushed the same day. On the next business day, wastewater was collected again and tested negative for SARS-CoV-2 viral RNA . This occurrence prompted a vaccination and testing campaign on 6 January 2022. Vaccine booster shots were administered to 117 workers, and nine workers were identified as positive for COVID-19 via antigen testing . Positive individuals were furloughed immediately. Although RNA concentration declined on the next sample dates), N1 was still detected with high concentrations . Subsequently, high RNA concentrations were detected on 1 February 2022 , prompting additional testing of 225 workers on February 3, among whom three were positive . One additional worker who was not tested on-site reported a positive test result the same day. Individuals were furloughed, and upon resampling of wastewater, RNA concentrations dramatically decreased to levels considered negative (Cq = 40.65) the next day. During subsequent sampling on 7 February 2022, SARS-CoV-2 RNA was undetected (During Omicron predominance, SARS-CoV-2 RNA was detected in wastewater on 11 distinct sampling dates. The date of 5 January 2022 marked one of the highest concentrations of N1 measured in wastewater at the agriculture facility (Cdetected .WBE efforts detected positive SARS-CoV-2 signals for 18 days at a food processing facility near the United States-Mexico border from November 2020 to March 2022. Detections occurred during Arizona’s second and third COVID-19 infection waves  Figure . YCEDA cConversely, negative wastewater results may not have represented actual SARS-CoV-2 infections, or lack thereof, in the facility. For example, the non-detect on 11 January 2021, where juxtaposed collection dates had positive samples, may have been caused by a lack of shedding from infected individuals or that those positive individuals did not defecate in the facility’s restrooms.In at least three cases, two during the Wild-Type and one during the Omicron, positive wastewater samples led to possible breaks in transmission and, therefore, potential outbreak aversions. In those three events, after wastewater was determined to be positive for SARS-CoV-2 RNA, and infection control guidelines were reinforced, clinical testing was performed: all employees were tested, and positive workers were identified and furloughed, thereby removing potential sources for onward transmission from those individuals. Interestingly, the utility of test-and-furloughing yielded different results based on the period of the COVID-19 pandemic. WBE and clinical testing diagnostic pairings were perhaps the most impactful during Wild-Type predominance when employee furloughing led to an immediate reduction in SARS-CoV-2 RNA in wastewater to undetectable levels. This observation may be due to the relaxation of early, stringent county-wide policies during the later phases of the pandemic, including mask-mandate policies .No employees reported illness, nor was wastewater positive during Alpha or Delta, which may be due to low case prevalence in the surrounding area . HoweverBoth during Wild-Type and Omicron predominance, SARS-CoV-2 RNA concentrations in the facility’s wastewater peaked and crested. However, our interpretations regarding transmission events shifted relative to the variant predominance. During Wild-Type predominance, when COVID-19 was novel and the scientific information regarding the virus was rapidly expanding, individuals adhered to masking practices and social distancing more rigorously both at work and in the community. Therefore, the transmission was more likely to have occurred during necessary congregation on shift. However, during Omicron predominance, “worry fatigue,” and COVID burnout were apparent , and altSeveral limitations are evident in this work which may compromise findings from both clinical and wastewater results. Regarding clinical testing, several documentation flaws were noted, including lost documentation regarding the exact number of individuals who were tested on January 6 and which specific COVID-19 antigen tests were used. Therefore, the individual diagnostic capabilities of each test may have biased diagnostic results . RegardiAs the pandemic progressed, we gained more familiarity and knowledge regarding how to use wastewater as a tool to institute public health action within the facility. During Wild-Type predominance, acquiring and deploying COVID-19 antigen tests was challenging, resulting in multiple instances where wastewater was positive with a lack of follow-up clinical testing . During The food processing facility never suspended production during the monitored period, whereas neighboring private businesses and schools were forced to shut down until outbreaks could be controlled—one business even closed permanently due to a COVID-19-related death. Consequently, this food processing facility could maintain the workforce and continue employing the hundreds of people who relied on this income while also serving the dozens of countries that purchased the company’s dates. The food processing facility attributes its success to several factors. First, a strong sense of community is shared among the facility’s staff. Staff refer to their coworkers as “a second family,” and many of the facility’s employees are related either biologically or through nuptials. This community perspective encouraged facility leadership to pursue WBE collaborations with YCEDA initially: to monitor the situation with real numbers and prevent their members from preventable illness. Workers were encouraged to contribute to the wastewater for accurate correlations between wastewater sample analysis and the presence of infection, and all staff understood the importance of testing and vaccination to prevent the spread of illness: the facility reports a >90% vaccination rate, at least 15% higher than Arizona’s ~74% vaccination rate . For theThis work is also novel through this intensive public-private partnership. Several studies have demonstrated the efficacy of WBE in monitoring and preventing COVID-19 outbreaks in aircraft , cruise This study has possible implications for border security between the United States and neighboring countries. Understanding the epidemiology of COVID-19 among individuals who reside in one country and work in a neighboring country may provide critical information for early warning, preparedness, and heightened awareness for possible surges in case numbers, especially during times when testing capacity may be burdened. Similarly, whole-genome sequence technologies may provide early warning to novel variant introductions. Such information should never be used to ostracize or scapegoat individuals and should be used solely for public health and disease prevention efforts for all individuals.In conclusion, WBE has applications in diverse settings, exemplified by this case study in a food processing facility near the Mexico-United States border. Wastewater monitoring for SARS-CoV-2 informed public health action overall—including infection control response and clinical testing. In combination with clinical testing, WBE potentially mitigated two outbreaks, demonstrating that when adopted by public health and private industry, WBE may prevent widespread illness and, thus, economic losses.


Most data on the transmission and outcomes of syphilis occur in community sexually transmitted infection (STI) clinics and have shown increased risk for other STIs in patients who test positive. There is limited data on outcomes in other lower risk populations that undergo universal syphilis screening, such as blood donors. This study describes the initial evaluation, follow-up, and STI risk of Air Force military basic trainees who screen-positive for syphilis at blood donation.Blood donor data was gathered from the Armed Services Blood Bank Center-San Antonio for Air Force basic trainees who were screen-positive for syphilis between January 2014- September 2021. For each screen-positive case, six screen-negative controls were analyzed. Prospective donors were unable to donate if they reported male-to-male sexual encounters, HIV positive partners, or a bacterial STI in the prior year. Electronic medical records of cases and controls were examined for demographic information, STI history, and additional STI positivity within three years.(Table 1). At time of screening, 3 (13%) trainees who screened positive for syphilis also tested positive for an additional STI. After initial treatment and screening, donors who screened positive for syphilis had a RR: 3.8  of developing an additional STI in the three years after blood donation compared to controls. Cases of N. gonorrhoeae (Table 2) in the 3-year period following blood donation were significantly higher in those who were syphilis screen-positive compared to those who were negative.A total of 63,375 basic military trainees donated blood at the Armed Services Blood Bank Center-San Antonio during the study period, of which 23  screened positive for syphilis There is limited data on follow-up of blood donors who screened positive for syphilis. This study found that in a low-risk population of military donors, those who screened positive for syphilis were at increased risk of future STIs compared to donors who were screen-negative. Syphilis screen-positive donors may be a potential target for future interventions to decrease the STIs burden in this population.All Authors: No reported disclosures.