For recipients with graft failure monthly costs begin to rise 3–4 months prior to failure, with incremental costs of over $38,000 during the month of failure. Mean annual total incremental costs of graft failure are over $150,000. Total costs post-transplant are strongly correlated with eGFR. Graft failure in the first year is an expensive, months-long process. Further reductions in early graft failures could yield significant human and economic benefits.Relationships between renal function and medical costs for deceased donor kidney transplant recipients are not fully quantified post-transplant. We describe these relationships with renal function measured by estimated glomerular filtration rate (eGFR) and graft failure. The United States Renal Data System identified adults receiving single-organ deceased donor kidneys 2012–2015. Inpatient, outpatient, other facility costs and eGFRs at discharge, 6 and 12 months were included. A time-history of costs was constructed for graft failures and monthly costs in the first year post-transplant were compared to those without failure. The cohort of 24,021 deceased donor recipients had a 2.4% graft failure rate in the first year. Total medical costs exhibit strong trends with eGFR. Recipients with 6-month eGFRs of 30–59 ml/min/1.73m  Kidney transplantation is the preferred treatment for end-stage kidney disease (ESKD) with greater patient survival than dialysis as well as far lower annual costs –3 Early This body of evidence has helped inform patient management practices aimed at maintaining renal function in the first months post-transplant to help realize the recent gains in long-term human and economic benefits , 22 ChieGraft failure is relatively rare in the first year post-transplant. However it is a very costly clinical event by itself in addition to the subsequent return to dialysis and, perhaps, later transplant. Overall, the economic costs of failure in terms comparing the costs of those with functioning grafts versus those on maintenance dialysis have been well-characterized over the years –27 For eThus, the time-dependence of the relationships between graft function and medical costs in the first year post-transplant are not well studied: what are the short-term economic implications of patient management practices that improve renal function in the first year post-transplant? Our aims were to describe relationships between total medical costs in the first year following deceased donor kidney transplant with overall renal function as measured by: 1) eGFR at different time points for recipients with graft survival of at least 12 months, and; 2) graft failure for those with survival ≤12 months. We used the USRDS claims dataset to assess these real-world economic relationships within 1 year post kidney transplantation.A retrospective cohort of single-organ, deceased donor kidney transplantation recipients was identified using the United States Renal Data System (USRDS) database from 1 January 2012 to 31 December 2015. One year follow-up was allowed to 31 December 2016. The USRDS database is a joint effort of the National Institute of Diabetes and Digestive and Kidney Disease (NIDDK) and the Centers for Medicare and Medicaid Services (CMS) that tracks many descriptive elements for all patients in the US with end-stage kidney disease (ESKD). USRDS registries integrate information from the Organ Procurement and Transplantation Network (OPTN), CMS, and Medicare billing claims records (This study population included adult (18+) deceased donor kidney transplant patients with Medicare as their primary payer at time of first transplantation from 2012 to 2015. If patients had multiple kidney transplant procedures in that time period, the first was included. Patients with other organ transplants prior to the index transplantation were excluded from this study.Patients without serum creatinine measures at discharge, at 180 and 365 days post-transplantation were excluded from this analysis . PatientThe following patient demographics and clinical variables were assessed in this study : age at Costs were derived from Parts A and B claims including inpatient, emergency, outpatient, and skilled nursing facility costs. Costs were included following discharge from the initial transplant hospitalization. Note that, for patients with graft failure, dialysis costs are not reported separately by USRDS and are largely incorporated into the Outpatient cost category. eGFRs were available at discharge, and at 6 and 12 months. Thus, eGFR:Cost relationships are described for several month-based time periods post-discharge: 0–3, 3–6, and 6–12 months. Serum creatinine measures were included in they were within 2 weeks of the discharge, 6, or 12 month time periods. All costs were adjusted to 2019 USD. In this study, eGFR was assessed using the Chronic Kidney Disease Epidemiology Collaboration (CKD-EPI) equation, using serum creatinine , 35.Descriptive statistics were used to describe the baseline demographic and clinical variables. Continuous variables were described with mean and standard deviation (SD). Categorical variables were described with counts and percentage. Data were missing for calculation of BMI for a few patients  and these missing values were imputed using the mean from patients with complete data. Both descriptive and multivariate generalized linear models with standard gamma distribution and log link function were used to define the relationship between eGFR measures and total medical costs.For recipients with graft failure a time-history of medical costs was constructed with failure as the index date (Month 0 in 2). A greater proportion of recipients with early graft failure were Black (40.8% vs. 36.3%) and were less likely to have PKD (3.2% vs. 6.8%) and more likely to have glomerulonephritis (24.2% vs. 21.3%) as cause of ESRD. Among those without graft failure in the first year post-transplant over 96% had serum creatinine measurements at discharge, and at 6 and 12 months post-transplant.Patients who received a single-organ deceased donor kidney transplant from 2012 to 2015 experienced a 1-year graft failure rate of 2.4% . CompareMean monthly medical costs in the first year post-transplant  are subsBy treatment setting, inpatient costs are the largest driver of cost differentials between patient groups. In the discharge to 3 month period, mean monthly inpatient costs are 4.4 times higher in the graft failure group  and 10.9 times higher  in the 6–12 month period. Outpatient costs are consistently higher for those with graft failure in the first year; during the early period mean monthly medical costs are 42% higher than non-failures  and, while absolute OP costs decrease overall, the differential rises to 81% higher during the 6–12 month period . Other medical costs  are small components of total costs but are consistently higher for those with graft failure.p < 0.001 for each trend). The trend in the discharge to 3 month time period appears to be the least strong likely due to high variance among serum creatinine values at discharge  and costs are another 35% lower for 30–44 ml/min/1.73 m2 as compared to 15–29 ml/min/1.73m2 . In addition to eGFR measures, age, gender, race, BMI, and cause of ESRD are important determinants of mean monthly costs in most time periods  and later time period (6–12 months), respectively. The peri-failure period is a time of particularly complex and intensive care management. Evidence that there is slightly higher patient mortality risk for each 1 ml/min/1.73 mitiation , 37 undeImportant limitations of this study are those typical for retrospective analyses of observational data, including the potential for residual confounding associated with factors not available in the study database. Additionally, given the study inclusion criteria, costs may not be generalizable to all DD recipient populations. For example, we excluded patients without Medicare as primary payer and our results may not apply to patients with employer group health plans. More generally, USRDS data lacks detailed laboratory and histology data that may be appropriate to include in our regression analyses of the eGFR:cost relationship regarding certain data elements and this lack of specificity may be important for prospective study. While additional control variables, if included, might be shown to be statistically significant determinants of first-year costs, our analysis suggests that they would not achieve the dominant role of renal function.Our findings demonstrate that total medical costs in the first year post-transplant are substantially correlated with renal function, measured both as graft failure and as eGFR among those with surviving grafts at various times post-discharge. These results indicate that continued focus on improving renal function and reducing early graft failures could yield significant human and economic benefits.


Medical textiles are subject to particularly harsh disinfection procedures in healthcare settings where exposure risks are high. This work demonstrates a fabric treatment consisting of a reactive silver ink and low surface energy PDMS polymer that provides for superhydrophobicity and antiviral properties against enveloped herpes simplex virus stocks even after extended ultrasonic bleach washing. The antiviral properties of reactive silver ink has not been previously reported or compared with silver nanoparticles. The fabric treatment exhibits high static contact angles and low contact angle hysteresis with water, even after 300 minutes of ultrasonic bleach washing. Similarly, after this bleach washing treatment, the fabric treatment shows reductions of infectious virus quantities by about 2 logs compared to controls for enveloped viruses. The use of silver ink provides for better antiviral efficacy and durability compared to silver nanoparticles due to the use of reactive ionic silver, which demonstrates more conformal coverage of fabric microfibers and better adhesion. This study provides insights for improving the wash durability of antiviral silver fabric treatments and demonstrates a bleach wash durable, repellent antiviral treatment for reusable, functional personal protective equipment applications. Personal protective equipment (PPE) such as gowns, masks, and scrubs are essential for protecting healthcare professionals from contact with bacteria and viruses that lead to infection. There is a need for textile coatings that provide PPE with better protection from infection by not only repelling fluids such as respiratory droplets, but also killing or deactivating microbes . In partEscherichia coli, and Staphylococcus aureus [There has been a tremendous amount of research on the use of silver nanoparticles as an antimicrobial agent , 6 and ts aureus . More res aureus –18.However, nanoparticles on fabrics may have issues with being removed in the wash, resulting in the loss of any associated functionality , 20. RecThere has been no previous work on the antiviral properties of silver fabric treatments after disinfection with bleach (sodium hypochlorite) concentrations as high as 10% and ultrasonic cleaning. Bleach (sodium hypochlorite) is a strong oxidant that is chemically reactive with organic molecules , 34. TheThis work studies the virucidal activity of single knit polyethylene terephthalate (PET) fabrics coated with a silver amine complex ink  against The reactive silver ink demonstrates better antiviral and bleach wash durability compared to 20 nm diameter silver nanoparticles. The size of the silver nanoparticles is considered as it is the maximum size to offer virus inhibition with HSV-1 and HIV-1 , 43. Alsin-situ on the textile and uniformly covers the microfibers of the fabric. This provides for better silver adhesion and uniformity with the fabric microfibers compared to nanoparticles, which adhere to the fabric with weak van der Waals forces. PET fabrics coated with 20 nm silver nanoparticles show a 1.1±0.6 log10  log reduction in virus quantities compared to control fabric, while PET fabrics coated with reactive silver inks reduce virus quantities by 2.3 ± 0.8 log10 .The chemical reduction of reactive silver ink into a pure silver film is previously described . The rea10  after 300 minutes of ultrasonic bleach washing. We demonstrate the combination of reactive silver ink with PDMS as a PET fabric treatment that adds durable, superhydrophobic and antiviral properties for reusable PPE applications.However, the results suggest the performance of both silver fabric treatments can be significantly hindered after extended periods of ultrasonic bleach washing. An additional polydimethylsiloxane (PDMS) thin film is used protect the silver layer and increase the liquid repellent properties of the fabric to further improve the bleach wash durability of the antiviral functionality. When the silver layer is protected by the polymer, the silver ink treated fabric retains 42% more silver than silver nanoparticle treated fabrics after bleach washing. PET fabric samples coated with reactive silver ink and PDMS reduce virus quantities by 1.7 ± 0.2 logθ = 38.2° and 44.5° corresponding to the (111) and (200) crystal planes. The XRD analysis confirms the presence of poly-crystalline on coated PET with silver nanoparticles  difference. Previous reports have suggested that virus inactivation is stronger with smaller diameter silver nanoparticles (≤ 15 nm) and it is possible the nanoparticles are too large to cover the binding sites of HSV-1 to inhibit cell entry [Additonally, we investigate the anti-viral activity of the silver coatings in solution . Virus aticles ≤  nm and iA PDMS post treatment is added to the silver coated samples to improve the ultrasonic bleach wash durability. PDMS is a low surface energy polymer that offers liquid repellent and chemical resistance properties to fabrics . Both PEθ = 22 − 27° due to the amorphous polymer structure.The cumulative percentage of silver that came off PDMS coated samples during ultrasonic bleach washing is shown in θ = 27.8°, 32.3° and 46.3° corresponding to the (111), (200) and (220) crystal planes, respectively, of silver chloride  on fabrics, using standard plaque forming unit (PFU) assays, for samples that are unwashed, bleach washed for 100 minutes and bleach washed for 300 minutes samples. Virus assays on fabric are conducted by submerging fabrics in virus/PBS while rocking for 1 hour, then samples are removed and placed in fresh PBS. Afterwards, samples are sonicated at low power to remove viruses that adhered on the samples. The PBS liquid afterwards is used to quantify the amount of virus present on the samples. Mann-Whitney tests are utilized to offer a degree of statistical certainty that the fabric treatment reduces the amount of virus particles on the fabric compared to controls. Asterisks corresponds to the level of certainty that the treated samples are the same as the controls where one asterisk corresponds to 10 , 2.3 ± 0.8 log10 , 2.3 ± 0.8 log10  and 2.8 ± 0.2 log10  PFU per mL, respectively. PET-Ink samples show less infectious virus than PET-NP samples likely due to the higher antiviral activity of the silver ink in liquid. Coating the samples with PDMS reduces the amount of infectious virus observed. PDMS swells in the presence of liquid which potentially allows silver to slowly diffuse [The amount of virus on fabric samples before and after ultrasonic bleach washing is shown in  diffuse , 46–48. 10 , 1.6 ± 0.4 log10 , 2.4 ± 0.3 log10  and 2.8 ± 0.1 log10  PFU per mL, respectively. Samples without the PDMS layer may lose some functionality as significant amounts of the silver is removed from washing. Samples with the PDMS layer maintain liquid repellency and silver after 100 minutes of bleach washing due to the chemical resistance of PDMS which retains the silver layer. The changes of the performance of the samples are not statistically significant after 100 minutes of bleach washing.The amount of virus on fabric samples after 100 minutes of bleach washing is shown in 10  PFU per mL due to the liquid repellency and virus inactivation properties. Overall, the PET-I/P treated fabrics reduce the amount of active virus, even after 300 minutes of bleach washing. The reactive silver ink coating covers the fibers more uniformly and the PDMS coating adds silver retention and liquid repellent properties for reusable, antiviral performance.Lastly, the amount of virus on fabric samples after 300 minutes of bleach washing is shown in There is a need for reusable, functional PPE to improve the public health safety against viral infections. This work demonstrates how reactive silver ink and a low surface energy polymer can be used to repel liquid and inactivate viruses on polyester fabric with lasting functionality under harsh ultrasonic bleach washing. The fabric treatment fully repels water before and after bleach washing. Most importantly, the fabric treatment shows reductions in enveloped virus quantities by about 2 logs compared to controls before and after bleach washing. Future work should analyze the antiviral mechanism, as well as the cytotoxicity of reactive silver ink treated fabrics before commercial use. All in all, this work demonstrates a durable, superhydrophobic, antiviral functionality on common fabric for reusable PPE applications.PET single knit cleanroom wipes (Anticon Gold), acetone (99.5%), methanol (99.9%) and isopropyl alcohol (99.5%) were bought from VWR. PBS (P5493), FBS (12103C), PDMS (Sylgard 184) and hexane were bought from Sigma-Aldrich. Reactive silver inks  was provided by Electroninks. Silver nanoparticles  were purchased from NanoComposix. Household bleach  was purchased from Giant Eagle. Deionized water was used from a Millipore Academic A10 system with total organic carbon below 40 ppb. Herpes simplex virus were obtained from a clinical isolate of HSV-1 Mckrae and frozen at -70°C. Virus stocks in PBS were prepared with A549 human lung carcinoma cells. The virus stocks were diluted in sterile PBS to the experimental titers used.0.5 in. by 0.5 in. square samples were cut from PET fabric. All samples were rinsed with acetone, methanol and isopropyl alcohol and dried with nitrogen to eliminate possible contaminants. An equal amount of 0.2 mg of silver was added to fabric samples, confirmed by microscale after curing. Silver ink was added by drop casting onto untreated PET fabrics followed by oven curing for 1 hour at 120°C to make PET-Ink samples. Silver nanoparticles was added by drop casting onto untreated PET fabrics followed by oven curing for 1 hour at 120°C to make PET-NP samples. Additional PET-Ink and PET-NP samples were treated with a PDMS layer by soaking samples in PDMS (Sylgard 184) at 1:10 curing agent:PDMS ratio dissolved in hexane and curing for two hours at 150°C to make PET-I/P and PET-NP/P samples, respectively.−1 using 16 scans.The physical morphologies of samples were characterized by scanning electron microscopy  at 5 kV. For SEM imaging, all samples were sputter coated with 10 nm gold/palladium (80:20) using a sputter coater (Denton). The presence of silver was confirmed using X-ray diffraction  locked coupled continuous scan at voltage 40 kV, current 40 mA and step size 0.04 degrees. The chemical compositions of samples were characterized by Fourier transform infrared spectroscopy  between wavenumber 600 and 3200 cmStatic, advancing and receding contact angle measurements were taken in ambient air at 22–25°C and 20–30% relative humidity using an optical tensiometer . 5 μL droplets at 25°C for all test liquids were used for all wetting measurements. The hysteresis was tabulated for each treatment after measuring the advancing and receding contact angles during syringe-controlled water dispersion and withdrawal, respectively.Breakthrough pressure was measured by observing the contact angle and volume while a water droplet evaporates. When the droplet transitioned from Cassie-Baxter to Wenzel state the diameter of the droplet was tabulated to calculate the breakthrough pressure.Washing cycles were performed using a Powersonic P230 Ultrasonic Cleaner (Crest) under ASTM G131–96 standards for washing materials by ultrasonic techniques. 10% bleach washing solution was prepared in a 10 mL test tube to create a highly efficient washing solution. Samples were submerged in bleach solution in Eppendorf tubes and ultrasonicated for 30 minutes at 80 W and 54°C to complete one wash cycle. Afterwards, samples were dried in ambient temperature before testing.Silver leaching analysis was conducted using inductively coupled plasma-mass spectrophotometer (ICP-MS). Wash affluent was collected as samples were subject to bleach washing cycles. Then, samples were diluted for ICP-MS by mixing 9.8 mL of water (2% nitrate solution), 10 μL of standard and 20 μL of wash affluent and taking the weights using a microscale. The weights were used to calculate dilution factors for ICP-MS results.2 and then fixed and stained with 0.5% gentian violet solution containing formalin and the plaques were quantified.Virus inactivation assays were performed by mixing silver nanoparticles or silver ink with herpes virus in PBS at final silver concentration of 1 mg per mL (0.1% v/v). Solutions were vortexed and incubated for 1 hour at room temperature. After 1 hour of incubation, 500 μL of ice-cold tissue culture media containing 20% FBS was added, then vortexed and centrifuged for 1 minute in the Eppendorf centrifuge to pellet any silver. The supernatants were removed and plated with 0.1 mL of serial 10-fold dilutions in duplicate onto A549 monolayers in 24 well multiplates. The virus was absorbed for 1–3 hours after which the wells were filled with 1 mL of tissue culture media containing 0.5% methyl cellulose. Plates were incubated for 5–6 days in 5% COFabric samples were completely submerged in 0.4 mL of virus/PBS in Eppendorf tubes with moderate shaking for 1 hour  at room temperature. After shaking, samples were gingerly rinsed once in sterile PBS and submerged in Eppendorf tubes with 0.4 mL of PBS. Then, virions attached on the surface were removed from the samples into PBS by sonication at power 3 for 10 seconds  within the tubes. The remaining PBS was used to quantify plaques.2, the cells were fixed and stained with gentian violet prepared in formalin, the number of plaques per well were counted under a dissecting microscope, and viral plaque forming unit titers were calculated. Finally, Mann-Whitney U-tests were performed at different p values for all comparisons between control and treated samples.Virus titers (PFU per mL) for herpes simplex virus were determined using standard plaque assay with A549 human lung carcinoma cells prepared in 24-well tissue culture plates. After 6–7 days incubation at 37°C in 5% COS1 File(DOCX)Click here for additional data file.S1 Graphical abstract(TIF)Click here for additional data file.


Mycoplasma genitalium, Mycoplasma hominis, Ureaplasma parvum, Ureaplasma urealyticum, Trichomonas vaginalis, and Gardnerella vaginalis, by retrospectively analyzing a cohort of patients living in the Naples metropolitan area, Campania region, Southern Italy. To investigate the genital infections prevalence in clinical specimens  collected from infertile asymptomatic women and men from November 2018 to December 2020, we used a multiplex real-time PCR assay. Of the 717 specimens collected, 302 (42.1%) resulted positive for at least one of the targets named above. Statistically significant differences in genital prevalence of selected microorganisms were detected in both women (62.91%) and men (37.08%). G. vaginalis and U. parvum represented the most common findings with an 80.2% and 16.9% prevalence in vaginal/endocervical swabs and first-voided urines, respectively. Prevalence of multiple infections was 18.18% and 8.19% in women and men, respectively. The most frequent association detected was the co-infection of G. vaginalis and U. parvum with 60% prevalence. Our epidemiological analysis suggests different infection patterns between genders, highlighting the need to implement a preventative screening strategy of genital infections to reduce the complications on reproductive organs.This study provides updated information on the prevalence and co-infections caused by genital microorganisms and pathogens:  Genital tract infections (GIs) remain one of the major global public health issues causing infertility, a condition that affects about 15% of couples of reproductive age, representing a globally underestimated problem with a significant psychological impact ,3.Chlamydia trachomatis, Ureaplasma spp., Neisseria gonorrhoeae, Trichomonas vaginalis, and Mycoplasma genitalium are the most common causative agents [C. trachomatis, N. gonorrhoeae, and T. vaginalis are classified as some of most common curable agents of sexually transmitted infections (STIs) [M. genitalium was recognized as an STI, albeit not a notifiable infection, and still few data exist on its prevalence [There are over 30 bacterial, viral, fungal, and parasitic pathogens known to contribute to infertility and among these human papillomavirus (HPV), e agents ,5,6. In s (STIs) ,8; recenevalence .GIs can cause a wide range of clinical presentations, both in the short and long term, such as urethritis, prostatitis, cervicitis, and vaginitis, which if unrecognized and left untreated may lead to relevant clinical complications including infertility, pelvic inflammatory disease, chronic pelvic pain, adverse pregnancy outcomes, premature delivery, neonatal death, and increased risk of contracting severe STI such as human immunodeficiency virus (HIV) infection ,14,15,16T. vaginalis infection, one of the most prevalent non-viral STIs, is often underestimated due to the high frequency of asymptomatic patients contributing to the dissemination of this infection [It is well known that the genital tract harbors numerous microorganisms defined as microbiota that exist (in conjunction with their genes and products) in a regulated mutualistic relationship with the host ,18. Somenfection .Several evidence show that a significant number of patients with an altered genital microbial balance that leads to GIs can result in co-infection with multiple pathogens at the same time ,21,22. FPrompt recognition and appropriate treatment are crucial for the control of GIs and their transmission and require sensitive, rapid, and accurate laboratory diagnostic assays. At the moment, the traditional methods for diagnosing GIs are laborious, with low sensitivity and a long turnaround time. Moreover, most of the organisms that cause GIs are difficult to detect with conventional microbiological strategies.In the past few years, new molecular tools have been developed in order to improve their diagnosis and to identify a wide array of microorganisms in any clinical sample at once.New real-time multiplex PCR (mRT-PCR) methods allow a rapid detection of genital pathogens from various specimens  and provide an additional advantage in screening since they detect multiple agents simultaneously from a single clinical sample with high sensitivity and specificity. Some of these specimens can be collected without the need of clinical examination, thus increasing their potential usefulness in screening strategies. However, some genital microorganisms are also common commensals in the urogenital tract of healthy people, but an increased bacterial load, evaluated by quantitative real-time PCR, can be considered an indicator of infection .Moreover, several recent reports showed that genital infection screening tests using multiplex PCR assays provided higher performances than the conventional methods ,25,26. TThe molecular screening could indeed reduce the risk of transmission to sexual partners leading to a reduction of treatment costs and a consequent decrease of antibiotic-resistant strains.U. urealyticum/U. parvum; M. hominis/M. genitalium, and T. vaginalis/G. vaginalis) simultaneously detected by mRT-PCR in different types of genital specimens in a cohort of patients undergoing fertility investigations at the University of Naples “Federico II” Hospital.The purpose of the current study was to investigate the prevalence and association of co-infections of six common genital pathogens  from the metropolitan area around Naples, Campania region, Southern Italy, were collected and processed by the Complex Operative Unit of Clinical Microbiology, University of Naples “Federico II” Hospital, Italy.In particular, specimens included vaginal/endocervical swabs from female patients and first-voided urines from male patients. Sample information  together with the data of patients for whom molecular testing was performed  were recorded in an anonymous database by changing sensitive data into alphanumeric codes. No clinical data associated with these specimens were available. Being a retrospective study, formal consent was not required.U. urealyticum/U. parvum; M. hominis/M. genitalium, and T. vaginalis/G. vaginalis, and were searched out both in first-voided urine and vaginal/endocervical swabs by multiplex real-time PCR. Vaginal/endocervical swab specimens were placed in 3 mL Amies Transport medium . The collection tubes and urine specimens were equilibrated at room temperature and mixed by vortexing, then 300 µL of samples were pre-treated with a tissue lysis buffer . DNA was extracted from the specimens using the instrument EZ1 Advanced XL (QIAGEN GmbH), according to the manufacturer’s instructions, and stored frozen at −20 °C until testing. The detection of mycoplasmas, ureaplasmas, and T. vaginalis/G. vaginalis was performed with the RealLine STI Pathogen Kits , commercially available mRT-PCR assays for the detection of genital pathogens DNA. The whole process was monitored adding to each sample 20 µL of Internal Control (IC) (BIORON Diagnostics GmbH), provided by the manufacturer, before the DNA extraction, to confirm the nucleic acid extraction and to exclude PCR inhibition. The mRT-PCR tests were performed according to the manufacturer’s protocol. Briefly, the amplification was performed in a CFX96 Real-Time thermocycler . Each PCR was performed with 50 µL of extracted DNA.The targeted genital pathogens, with fastidious growth requirements or non-cultivable, were The thermal cycling conditions consisted of an initial incubation at 50 °C for 2 min, pre-denaturation at 95 °C for 2 min, followed by 50 cycles of alternating incubations: denaturation at 94 °C for 10 s, and annealing and extension at 60 °C for 40 s.G. vaginalis and U. parvum which were considered positive with a Ct value of ≤32, because these bacteria are normally part of the genital flora of healthy people and the positivity to the test should reflect their overgrowth more often associated with genital discharge and dysbiosis.Samples were considered positive with an average Ct value of ≤40, except for p < 0.05.A descriptive analysis was performed to assess the main characteristics of all the subjects enrolled in this study . Considering the characteristics of the variables and sample size, Chi-Square Test and Fisher’s exact non-parametric tests were used to test hypotheses about the relationship between two variables and to test differences between proportions for two populations. To measure an association with exposure, we computed the odds ratio by the classical approach (logistic regression). All tests performed were considered significant for a value of Between November 2018 and December 2020, a total of 717 subjects from the metropolitan area around Naples, Campania region in Southern Italy, were recruited for the study. Subjects must not have used antibiotics in the 15 days prior to the analysis and must not have had any type of sexual intercourse during the 48 h preceding the sample collection.The general demographic characteristics of the enrolled subjects are presented in p-value < 0.001), conversely most of the evaluated men were negative for the pathogens’ detection . Amongst the positive genital pathogen detection tests (302), 190 women tested were positive with a percentage of 62.91% (190/302) compared to 112 men who tested positive with an incidence of 37.08% (112/302)  .The retrospective analysis involved mainly patients aged between 30 and 39 (362 patients), followed by subjects ranging from 40 to 49 years (232), while the lowest number of patients were over 50 years old (55) .p-value < 0.001) and over 50    than subjects under 30 and over 50   . Patient < 0.05) . Among t over 50 . Moreove < 0.05) .Indeed, the risk of GIs in the age group 30–39 years was higher : 1.66, CI: 1.23–2.24) than the other age groups considered in our study. In detail, almost 50% of detected pathogens affected this age group , with a The distribution of single and multiple infections for one or more genital microorganisms tested by mRT-PCR analysis is shown in G. vaginalis and U. parvum represented the most common findings with a prevalence of 80.2% (166/207) and 16.9% (35/207) respectively , M. genitalium  and T. vaginalis   , G. vagiectively A. Other ; 0.48%) A.G. vaginalis and U. parvum  resulted the most prevalent multiple infection, followed by G. vaginalis and U. urealyticum   B; the re95, 15%) B. Triple95, 15%) B. Among inations B.With regards to the pathogen distribution according to gender, Our results showed that 52.34% (190/363) of women and 31.63% (112/354) of men were positive for at least one of the six genital microorganisms detected by mRT-PCR .G. vaginalis DNA, 27/363 (7.43%) for U. parvum DNA, and 1/363 (0.27%) for U. urealyticum DNA, respectively  and U. parvum DNA in 8/354 (2.25%) .Overall, among vaginal/endocervical swabs collected from women, 96/363 (26.44%) were positive for ectively . With re (2.25%) . For U. In the case of multiple infections, the prevalence was 18.18% (66/363) and 8.19% (29/354) in samples collected from women and men, respectively , indicatU. urealyticum was found predominantly in multiple infections, regardless of gender, whereas, G. vaginalis was the prevailing agent of single infections , T. vaginalis (0.48%), and M. genitalium (0.97%).Although the prevalence of genital pathogens shown in this report was greater among women (62.91%) compared to men (37.08%), G. vaginalis and U. parvum (60%). Globally, female patients were significantly more affected than male patients in both single and multiple infections.The evaluation of microbial co-infection prevalence showed that the most frequent associations detected in women and men were due to Considering the age of the analyzed subjects, the risk of GIs in the age group 30–39 years was higher than in the other age groups examined in our study, with a higher risk for both single and double infections.G. vaginalis and U. parvum were the most detected microorganisms in both women and men.In accordance with other published studies ,33,34, oG. vaginalis was found in 40% of healthy women. Even if its role is yet to be fully elucidated, virulent strains of G. vaginalis are able to create microbial alterations by inducing bacterial vaginosis through adhesion to vaginal epithelial cells and biofilm formation [G. vaginalis could also be associated to cervicitis, endometritis, salpingitis, and other relevant clinical implications [ormation ,36. MoreUreaplasma spp. were predominantly identified in the case of multiple infections. Although Ureaplasma spp. are considered part of the normal genital flora with an average colonization rate of 40–80% [U. parvum to form biofilm favoring its persistence, which contributes to the pathogenesis of chronic infections and facilitates mixed infection with other bacteria [Ureaplasma spp. in bacterial vaginosis remains highly controversial, since their presence has been demonstrated in a large proportion of women both with (65%) and without (48%) bacterial vaginosis [Ureaplasma spp. Among the most notable is the role of Ureaplasma spp. in adverse pregnancy outcomes, such as chorioamnionitis and premature rupture of membranes leading to preterm birth [Ureaplasma and G. vaginalis represents a potential impairing factor of fertility [f 40–80% ,38 they f 40–80% . An in vaginosis . Numerourm birth ,43,44. Fertility .It is well documented that some of the discussed microorganisms may be present as commensal flora in most of the sexually active people; due to their colonizing role, it is difficult to elucidate the pathogenic involvement of these agents, suggesting the need for epidemiological studies.Although many efforts have been made to better diagnose the infections of genital tract in patients with symptoms , multiplFor these reasons, further studies are needed in order to develop new GI control strategies. The subjects enrolled in the present study were predominantly asymptomatic and had no personal STI history. If sexually active women and men with or without symptoms could be tested for the screening and diagnosis of GIs by multiplex molecular assays, it might be helpful to simultaneously detect several microorganisms.G. vaginalis and the observed association between G. vaginalis and U. parvum highlighted the importance of molecular screening as a valid tool for identifying infections in sexually active people which can be treated with the aim of reducing the risk of clinical complications.In the present study, the significant presence of U. urealyticum/U. parvum; M. hominis/M. genitalium, and T. vaginalis/G. vaginalis) among a large asymptomatic female and male population in a selected area of the Campania region in Southern Italy.In conclusion, our study provides an estimation of the prevalence of single and multiple infections caused by six common genital pathogens (Overall, our epidemiological analysis reveals different patterns of infection between the genders and could help to identify groups at higher risk for clinical implications of GIs, underlining the need to implement a preventative screening strategy of genital pathogens.


BackgroundPosterior tibial slope (PTS) is one of the factors that determine the postoperative range of movement (ROM) in total knee replacement (TKR). While biomechanical factors influencing ROM such as PTS, soft-tissue balancing, and choice of implants are surgeon-dependent, non-biomechanical factors such as physiotherapy and pain are subjective and beyond the surgeon’s control. Using navigation, we avoided these factors and objectively correlated the difference in PTS with ROM.MethodologyA total of 120 cases of posteriorly stabilized (PS) TKR were included. The X-ray-measured difference in PTS was compared to the corresponding change in the ROM intraoperatively using the TKR navigation system. Based on the change in PTS, the cases were classified into three groups, and the intergroup variability of mean postoperative ROM, mediolateral laxity difference (MLD), and residual flexion (RF) was calculated.ResultsAn average MLD of 1.39 mm in extension and 1.79 mm in flexion and an average RF of 3.18 degrees were seen. This uniformity neutralizes the effects of these factors on the ROM. The mean difference in the PTS in the three groups was 4.45, 10.76, and 17.98 degrees, and the mean change in the ROM was 3.07, 1.47, and 2.84 degrees, respectively. There was no statistically significant change in ROM with that of change in PTS.ConclusionsIn our study, it was shown statistically that the coronal and sagittal planes were uniformly balanced using navigation assistance, and the PTS and the postoperative ROM were the only variables. The correlated results showed that a change in the PTS does not affect the postoperative ROM in PS TKR using the implant system used in our study. Total knee replacement (TKR) is a well-accepted procedure for the management of advanced arthritis of the knee, providing pain relief and adequate knee movements to perform daily activities . Hence, Among the various biomechanical factors, the contribution of PTS in determining ROM remains a subject of debate. Some studies have shown a positive correlation between the two variables ,7, whereBased on the results of a few earlier studies on ROM in TKR ,7, we hyA total of 120 TKRs in 96 patients performed at our center using navigation assistance were included in the analysis. The study included cases from March 2018 to March 2019. Posteriorly stabilized (PS) rotating platform implants  and computer navigation using the Brainlabs™ system were used for all cases. Knees with gross bony irregularities, defects, or extra-articular deformities where precise radiological measurements were not possible were excluded. An institutional and ethical review board approval was not considered as the study is a retrospective observational analysis of the data routinely recorded while performing the navigation-assisted TKR.The preoperative and postoperative PTS were measured from the digital radiographs by two orthopedic surgeons, and the average of the two values was taken for the analysis. Lateral views of the knees were taken in such a manner that both the femoral condyles superimposed each other and had 15 cm of proximal tibia included in the film. Two lines were drawn as follows: line 1 was drawn by connecting the midpoints of the outer cortical diameter at 5 cm and 15 cm distal to the knee joint, and line 2 was drawn tangential to the tibial plateau in the preoperative radiographs and parallel to the tibial base plate in the postoperative radiographs . The acuAll cases were operated on by the same surgeon (supervising author). After the placement of navigation arrays over the distal femur and proximal tibia, the hip center point, the distal femur mechanical axis point, the proximal tibial mechanical axis point, and the lateral and medial malleolar points were registered. The free maximum flexion was noted using navigation before the surgery by lifting the thigh vertically and allowing the knee to flex freely under gravity. TKR was then performed using the navigation technique as per the Brainlabs™system. As the implants used were of rotating platform type, it was aimed at creating a slope close to 0 degrees  with theAt the end of implantation, the medial and lateral laxities were numerically assessed by monitoring the medial and lateral gaps from the navigation monitor after giving valgus and varus stress, respectively, to the knee in full extension and in 90-degree flexion. In all cases, balancing was done in the recommended manner using navigation for the implant in such a way that the difference between medial and lateral laxities (MLD) after implantation was within 1-3 mm. This was later statistically analyzed to check for uniformity among cases to make difference in PTS and difference in ROM as the variables. Similarly, residual flexion (RF) at the end of implantation was aimed at about 3-5 degrees for all cases as per the recommendations of the navigation system. This could indicate that at the end of implantation, the tissue tension was maintained uniformly, which may otherwise affect the knee ROM. After final implantation, the postoperative maximum free flexion was routinely recorded in the same manner which was used to record the preoperative ROM.Statistical analysisThe observed values were tabulated, coded, and analyzed using SPSS, version 17 for Microsoft Windows . Pearson correlation between the reduction in the PTS and the difference between the preoperative and postoperative knee ROM was calculated.The cases were divided into three groups  based on the difference between preoperative and postoperative PTS. Postoperative flexion, postoperative RF, and the mediolateral laxity difference (MLD) at the end of implantation were compared among the three groups using analysis of variance. P-values of <0.05 were considered statistically significant for all analyses.A total of 120 patients were included in the analysis. The respective preoperative and postoperative mean PTS, ROM, and the difference between the preoperative and postoperative values are listed in Table A scatter plot based on our analysis Figure  showed tThe MLD was 1.39 mm for extension and 1.79 mm for 90° flexion. There was no statistically significant deviation of this value from the mean for all cases. Similarly, the RF at the end of implantation was aimed at 0-5° for all cases. The mean RF obtained was 3.18°, and the cases had no significant deviation from the mean value.The change in the PTS from the preoperative to the postoperative values did not show any statistically significant corresponding change in the ROM measurements Table .Using Pearson correlation, the p-value for the analysis was -0.110. There was no statistically significant variability of the analyzed parameters  among the three groups, implying that these factors were uniform among cases, which might otherwise skew the ROM values. The corresponding analysis is presented in Table Postoperative ROM is an important factor that determines the success of TKR. Among the variables that determine postoperative ROM, PTS has been considered to be one of the most biomechanically important factors . Some stAnother important factor to be considered in our analysis is the mode of ROM measurements. Some of the commonly used modalities include the use of clinical goniometers, radiographic measurements, and, more recently, the use of mobile phone applications ,15. MeasFor measuring PTS, we used preoperative and postoperative lateral radiographs. Various methods of measurement have been suggested by various authors in a true lateral radiograph of the knee. We followed the already mentioned method, as suggested by Yoo et al. . Even thThe mean preoperative PTS in our study was 12.76 degrees. Mohanty et al.  reportedIn our study, the average preoperative and postoperative flexion angles measured using computer navigation were 119.80 and 117.79 degrees, respectively. On analyzing the difference in PTS with the difference in flexion angle, no statistical significance was noted p = -0.33). Our results correlated with Kansara et al. who also reported no significant relationship between the PTS and ROM in PS TKR 3. Our re,23 were Our study has certain limitations. First, we performed the retrospective analysis using only a single implant manufacturer (Depuy™). Therefore, the results of this study may not represent TKR performed using other systems. Second, regarding the method of performing TKR, all cases were performed using the PS method; hence, the results of the study cannot necessarily apply to the CR method.The findings of our study conclude that PTS has no influence on the ROM in PS TKR using the included implant. Our study also proposes navigation assistance as a method to record the ROM difference by negating the non-biomechanical factors that affect the postoperative ROM.


Heavy menstrual bleeding (HMB) is one of the most common distressing complications of uterine fibroids (UF); however, data on the health care costs for treatments in women experiencing HMB associated with UF are lacking. The objective of this study was to compare the direct costs and treatments patterns for women diagnosed with UF+HMB, UF only, and HMB only in the United States.® Commercial Claims and Encounters Database  and matched by demographics and Charlson Comorbidity Index score. Baseline characteristics and treatments during the 12 months post-diagnosis were summarized descriptively. Costs (2018 U.S. dollars) during the post-diagnosis year were compared using analysis of variance.The study design was retrospective matched cohort study using claims data. Women, aged 18–51 years, comprising four cohorts  were identified in the IBM MarketScanp < 0.001). The mean diagnosis-related costs were significantly higher for UF+HMB  than for UF only  and HMB only  . Surgery/procedure costs comprised 80% of diagnosis-related medical costs for UF+HMB.Before matching, women with UF+HMB represented 54% of UF cases. Following diagnosis, 32% in the matched UF+HMB cohort had no treatment, 49% underwent surgeries/procedures with (12%) or without (37%) medications, and 18% received medications only. The mean all-cause total costs for UF+HMB  exceeded that for UF only by 24%  and HMB only by 50% , and almost tripled the mean cost for the control cohort  (all, UF with HMB were associated with significant economic burden, driven primarily by surgical/procedural costs and treatment patterns. A 2012 review of published cost estimates cited annual direct and indirect costs ranging from $5.9 to $34.4 billion for women seeking treatment for UF in the United States.5 UF is the primary cause of hospitalization related to a gynecological disorder6 and accounts for up to 50% of hysterectomies.7–9 A retrospective cohort study in the United States also implicated UF as the cause of bleeding in nearly half (48%) of hospitalized women discharged with HMB and extremely low hemoglobin levels.10Uterine fibroids (UF), also known as leiomyomas, represent the most common benign gynecological tumors.11–13 Most studies have focused on the costs for surgery or inpatient care for UF, primary cost drivers in this population; 13 however, none have examined costs in the context of treatment practices or the absence or presence of HMB. To address this gap in real-world evidence, this study employed a large administrative claims database to examine the health care costs and treatment patterns for mutually exclusive, matched cohorts of women diagnosed with UF only, HMB only, or both UF and HMB (“UF+HMB”), and neither diagnosis. The primary aim was to estimate the direct health care costs and treatment patterns associated with these distinct clinical conditions while assessing the respective cost burden to the U.S. health care system.Numerous studies have quantified the direct costs of UF in the United States using different methods and cost measures.® Commercial Claims and Encounters Database  for the period October 1, 2007, through September 30, 2018. MarketScan provides electronic claims data submitted by large employers, managed care organizations, and hospitals in the United States for more than 160 million covered lives. Cases and UF/HMB diagnosis-specific claims were identified by International Classification of Diseases, Ninth (“ICD-9”) and Tenth (“ICD-10”) Revision, Clinical Modification codes. Prescription drug claims were identified by National Drug Codes and Healthcare Common Procedure Coding System codes. All database records are statistically de-identified and certified to be fully compliant with U.S. patient confidentiality requirements set forth in the Health Insurance Portability and Accountability Act of 1996. Because this study used only de-identified patient records and did not involve the collection, use, or transmittal of individually identifiable data, institutional review board approval to conduct this study was not necessary.This retrospective cohort study analyzed data from the IBM MarketScanFour cohorts were considered eligible based on diagnosis: (1) UF only, (2) HMB only, (3) UF+HMB, and (4) a control cohort with neither diagnosis. Eligible women in the UF only cohort had ≥1 diagnosis of UF (ICD-9 218.x or ICD-10 D25.x) but no diagnosis of HMB ; those in the HMB only cohort had ≥1 diagnosis for HMB, but none for UF. Women in the UF+HMB cohort had diagnoses for both UF and HMB, whereas those eligible for the control cohort had neither diagnosis during the study period. The index date represented the date of first UF diagnosis for women with UF only or the date of the first HMB diagnosis for women with HMB only. For the UF+HMB cohort, the index date was the later date of first diagnosis of UF or HMB. Women in the control cohort were assigned random index dates from a uniform distribution within the same period.Eligible women were included if they were aged 18–51 years on the index date and continuously enrolled for ≥1 year before the index date (“pre-index” baseline period) and ≥1 year following the index date (“post-index” follow-up). Women who had a diagnosis of gynecological cancer during the study period or a pre-index history of hysterectomy, myomectomy, uterine artery embolization, endometrial ablation, or magnetic resonance-guided high-intensity focused ultrasound were excluded.14 The primary outcome was direct all-cause and diagnosis-related health care costs incurred during the 1-year post-index period. Total costs were estimated as the sum of medical  plus pharmacy costs. “Other costs” pertained to costs for health services rendered in a residential/patient home, rural health clinic, urgent care facility, independent clinic, or unlisted facility. All costs represented gross payments to providers adjusted to 2018 U.S. dollars using the Consumer Price Index. Diagnosis-related costs only included those which had at least one UF or HMB diagnostic codes associated with the same observation.Outcomes were analyzed in the four cohorts of women matched 1:1:1:1 by age, geographical region of residence, insurance plan, and Charlson Comorbidity Index (CCI) scores.Treatments analyzed focused on either surgeries and procedures  or medications. Medication use included contraceptives, gonadotropin-releasing hormone agents, non-steroidal anti-inflammatory drugs, or antifibrinolytic agents.Patient demographics as of the index date and the CCI and the frequency of anemia diagnoses during the 1-year pre-index period before matching were reported. Outcomes analyzed in the matched cohorts included the pre- and post-index frequencies of anemia diagnoses, contraceptive use, inpatient admissions, and treatments .n and proportion) for categorical variables. Analysis of variance (ANOVA) was used to compare the 1-year post-index costs among the four matched cohorts. A p-value <0.05 indicated statistical significance.Statistical analyses were conducted using SAS, Version 9.4 . Patient demographics, clinical attributes, treatment utilization and patterns, and costs are summarized descriptively as the mean ± standard deviation for continuous variables or frequency  and nearly one third (31%) of those with HMB . Women with UF, regardless of a diagnosis of HMB, were on average older. More than 70% were aged ≥39–51 years in the UF+HMB and UF only cohorts compared with less than 50% in the HMB only and control cohorts.Before matching, the study sample consisted of 3,377,957 women , including 237,070 (7%) with UF only, 623,810 (18%) with HMB only, 280,636 (8%) with UF+HMB, and 2,236,441 (66%) with neither diagnosis . Women dA total of 836,992 women —209,248 from each cohort—were matched and included in the analyses. With the exception of the control cohort, the frequencies of anemia among the cohorts were higher during the post-index period. The UF+HMB cohort featured the highest proportions diagnosed with anemia during both the pre-index and post-index periods , followed by the HMB only , UF only , and control  cohorts.During the pre-index year, the majority in the matched cohorts with UF only (78%), HMB only (79%), and UF+HMB (72%) had received no UF/HMB-related treatments. Among treated women, those diagnosed with UF+HMB used contraceptives and other medications more often  than women with UF only  and HMB only . During the post-index year, UF/HMB-related treatments were recorded in 67%, 51%, and 38% of women in the UF+HMB, HMB only, and UF cohorts, respectively . Nearly n = 52,270) tended to be older than those who received myomectomies ; 81% in the former cohort were aged 39–51 years, whereas the majority (59%) in the latter were in their late 20s or 30s. Utilization of post-index surgeries/procedures among women with anemia appeared similar to the utilization in the overall study cohorts without anemia during baseline.Examining treatment patterns among the respective subgroups with (1) no treatments and (2) contraceptive use before diagnosis , treatmep < 0.001) (p < 0.0001).Approximately 5% of women in the matched cohorts had ≥1 inpatient admission during the pre-index period. During the post-index year, the UF+HMB cohort experienced the highest frequency of inpatient admissions (22%), followed by the cohorts with UF only (18%), HMB only (7%), and neither diagnosis (5%). The mean all-cause total cost for the UF+HMB cohort  exceeded that for the cohorts with UF only  and HMB only  by 24% and 50%, respectively, and was more than twice the mean cost for the control cohort    and SAT2p < 0.0001).Over half (59%) of the medical costs incurred by women with UF+HMB were diagnosis-related compared with 39% and 33% of women with UF only and HMB only, respectively. Diagnosis-related inpatient and outpatient costs accounted for higher proportions of the respective all-cause component costs in the UF+HMB cohort  than in the UF only  and HMB only  cohorts. Similarly, UF/HMB-related surgery/procedure costs represented higher percentages of the total all-cause and diagnosis-related medical costs in the UF+HMB cohort  compared with the cohorts with UF only  and HMB only  . The meapost hoc Tukey honestly significant difference pairwise comparisons showed that women with neither diagnosis on average incurred significantly higher pharmacy costs compared with the other cohorts , whereas women diagnosed with UF only had significantly higher mean pharmacy costs than those with UF+HMB .Pharmacy costs comprised a larger proportion of total health care costs in the control cohort (26%) than in the cohorts with HMB only (17%), UF only (15%), and UF+HMB (11%). ANOVA with 15 When clinical manifestations occur, HMB is the most common and often troubling symptom that can bear debilitating effects on health, daily activities, and quality of life.4 While women with UF who experience HMB may be expected to carry a higher disease burden than asymptomatic cases, data on the incremental cost impact are lacking. To the best of our knowledge, this is the first study to examine the direct health care costs and treatment patterns of women in the United States who were diagnosed with either UF or HMB or both conditions.Asymptomatic UF have been reported in 50%–75% of cases and may be detected incidentally during routine examinations or pelvic imaging.16 Among the cohorts, anemia was most prevalent among the women with UF+HMB. These cases exhibited nearly twice the pre- and post-index rates of anemia compared with women who experienced HMB without a diagnosis of UF. Compared with those with UF only, women with UF+HMB incurred excess mean total costs of 24%  during the post-index year. A prior claims analysis of the MarketScan database (January 2010‒December 2014) cited all-cause total surgical costs  averaging $15,813 ± $13,804 (2017 USD) for 95,433 commercially insured women with UF .13 Although the analysis did not differentiate between the costs for UF cases with and without a HMB diagnosis, the mean total cost appeared to approximate the estimated mean total health care costs for women with UF+HMB and UF only observed in the present study.In the study population of nearly 3.4 million women of reproductive age, the cumulative incidence of UF diagnosed over the 11-year period was 15%. HMB was diagnosed in over half (54%) of these cases. Overall, the UF+HMB cohort represented an older population; nearly 80% of women were aged ≥39–51 years. In contrast, women aged ≥39–51 years account for 36% of women aged 18–51 years in the general U.S. population.11 UF/HMB-related costs were primarily driven by the utilization of surgeries/procedures, particularly hysterectomy, and represented as much as 80% of the diagnosis-related medical costs for UF+HMB. About half of the women with UF+HMB treated with contraceptives before diagnosis underwent ≥1 surgery/procedure during the post-index year, suggesting heavier symptomatology or the need for a more effective management approach. Despite lower utilization of surgeries/procedures in the UF only (18%) and HMB only (24%) cohorts , the associated costs accounted for the majority (61% and 72%) of diagnosis-related costs.As previously reported,19 while the post-index year generally represents the peak period for pharmacological treatments for women managed nonsurgically.20 Examination of pre- and post-index treatment utilization revealed that the use of UF/HMB treatments increased following diagnosis, but only a modest proportion (38%) with UF only were treated. Some UF cases may have been asymptomatic and followed via an expectant management approach.15 The underlying reasons, however, could not be ascertained from the claims data. Other factors, such as patient preferences and behaviors in seeking medical care, fertility concerns, and variations in clinical practice, treatment options, and access to women's health care specialists, may have also influenced treatment decisions21 and merit future investigation.A majority of women with UF undergo ≥1 surgical or diagnostic procedure in the year following diagnosis,p < 0.0001). This finding appeared consistent with a prior study demonstrating a 68% increase in the mean costs for women with HMB versus controls .18Including the HMB only cohort allowed examination of cost differences between UF with HMB and HMB due to other causes. Overall, treatment for HMB only was less costly than UF regardless of a concomitant diagnosis of HMB. This may be attributable to the comparatively higher mean costs for inpatient admissions and surgeries/procedures in the UF cohorts. Compared with the control cohort, however, managing women with HMB only increased average total health care costs in the post-index year by 66% . Identification of the cohorts, costs, and resource use also relied on diagnostic codes rather than rigorous formal assessments and may be subject to errors. Additionally, there are no diagnostic codes currently dedicated to HMB associated with UF, and therefore, a recorded diagnosis of HMB did not necessarily indicate HMB caused by UF. Furthermore, while costs were compared between matched cohorts, the claims data were not sufficiently detailed to control for all factors potentially affecting treatments and costs. For example, several studies have reported that black women are significantly more likely than white women to be diagnosed with UF at a younger age and experience more severe symptoms.23 and incur costs for symptomatic treatment. Additionally, the study analyzed only pharmaceutical and surgical/procedural interventions, whereas previous literature has shown that lifestyle factors, such as physical activity, dietary changes, and weight management, can mitigate the risk of UF24 and may bear cost implications.24 While the follow-up period was limited to 1 year, some studies suggest that the likelihood of hysterectomy among women with UF may increase over time.25Other factors may have contributed to the underestimation of costs, but nevertheless have implications for future research. The retrospective claims-based nature of this study precluded consideration of undiagnosed UF cases, which may be prevalentStrengths of this study include that the IBM MarketScan Commercial Claims and Encounters Database was used as the source for the patient electronic health records, which is a nationwide database that allowed us to include records from individuals of various geographical locations in the United States and races and ethnicities. Another advantage of this database is that it includes claims for mail order prescriptions and specialty drugs, such as injectables.UF complicated by HMB were associated with significantly higher direct health care costs compared with UF or HMB alone. While surgical and procedural costs contribute substantially to diagnosis-related medical costs, effective pharmacological strategies can alleviate the clinical burden of symptomatic UF, particularly HMB, by reducing surgeries and procedures, thus ameliorating associated health care costs.


Genetic variants of vaccine poliovirus type 2, imported from an unknown source, were detected in waste waters in Jerusalem, London and New York in early 2022. Wild poliovirus type 2 was globally eradicated in 1999, but vaccine virus type 2 continued for 16 more years; routine use of the vaccine was discontinued in 2016 and reintroduced occasionally on purpose. As an unintended consequence, type 2 vaccine virus variants  that mimic wild viruses’ contagiousness and neurovirulence, have been emerging and spreading. To illustrate, in just the past four years (2018–2021), 2296 children developed cVDPV polio in 35 low-income countries. Many assume that virus transmission is via the faecal–oral route. Sustained virus transmission was documented in London and New York, in spite of high standards of sanitation and hygiene. Here, virus transmission cannot be attributed to faecal contamination of food or drinking water . Hence, contagious transmission can only be explained by inhalation of droplets/aerosol containing virus shed in pharyngeal fluids (respiratory transmission), as was the classical teaching of polio epidemiology. If transmission efficiency of VDPV is via the respiratory route where hygiene is good, it stands to reason that it is the same case in countries with poor hygiene, since poor hygiene cannot be a barrier against respiratory transmission. By extrapolation, the extreme transmission efficiency of wild polioviruses must also have been due to their ability to exploit respiratory route transmission. These lessons have implications for global polio eradication. It was as a result of assuming faecal–oral transmission that eradication was attempted with live attenuated oral polio vaccine (OPV), ignoring its safety problems and very low efficacy in low-income countries. Inactivated poliovirus vaccine (IPV) is completely safe and highly efficacious in protecting children against polio, with just three routine doses. Protecting all children from polio must be the interim goal of eradication, until poliovirus circulation dies out under sustained immunisation pressure. OPV should be discontinued under cover of immunity induced by IPV to stop the emergence of new lineages of VDPVs, not only type 2, but also types 1 and 3, to expedite the completion of polio eradication. The Global Polio Eradication Initiative (GPEI) announced on 29 July 2022 the recent detection of type 2 poliovirus  in sewage in Jerusalem, London and New York . One uniWild poliovirus (WPV) type 2 was eradicated in 1999 . Sabin-2Against this background, importations of variants of Sabin-2 virus in 2022, attest to the flawed immunisation tactics of the GPEI, particularly in using the vaccine for 16 years beyond its legitimate need, and also reintroducing it even after its global withdrawal.In recent years, only inactivated poliovirus vaccine (IPV) has been used in the UK (since 2004) and USA (since 1999). News media have suggested that the reason for reintroduction and sustained transmission of poliovirus is the exclusive use of IPV that does not induce intestinal immunity . HoweverThe silver lining of the dark clouds of these unfortunate events is that cVDPV-2 in low-income countries, from where it must have been imported, has now received global attention. There, 334 children were paralysed by cVDPV-2 in 2021 and 85 in 2022 (till 18 October) [The poliovirus genome is single-stranded positive sense RNA, highly prone to mutations. After many centuries of human adaptation, wild poliovirus (WPV) types 1, 2 and 3 have relatively stable genes for structural proteins. WPVs are highly contagious and neurovirulent, the two properties Albert Sabin managed to attenuate through repeatedly selecting and propagating variants with reduced virulence in monkeys .Thus, OPV contains wild-virus-derived polioviruses. During every cycle of multiplication, either in cell culture or human intestines, Sabin original (SO) virus mutates in the direction of the parental WPV genotype. Neurovirulence is regained after very few mutations. OPV is manufactured using SO passaged twice (SO+2) or thrice (SO+3) as seed virus, and next harvest as the final product. As a result, the presence of mutated neurovirulent virus particles in very small amounts in every dose of OPV is inevitable ,11.Currently, the entire VP-1 region of poliovirus genome of all field isolates is amplified and compared with that of the virus in OPV, designated ‘Sabin virus’, to distinguish variants drifted further from it. Genetic drift up to 0.5% for type 2 and up to 0.9% for types 1 and 3 are the criteria to classify variants as ‘Sabin-like’ (SL) . The virGenetic drift of 0.6% and more for type 2 and 1% or more for types 1 or 3 denotes that they are vaccine-derived polioviruses (VDPVs) that have regained neurovirulence and contagiousness. When epidemiological evidence shows community circulation of a VDPV, it is designated cVDPV, which is nearly fully deattenuated, and wild-like. About 1% drift denotes approximately one year of community circulation—as if a ‘molecular clock’ exists.In Egypt, Sabin-2 virus with genetic drift circulated from 1983 till 1993, causing polio, until stopped with tOPV campaigns—the drift from Sabin-2 virus was about 10% in ten years, validating the molecular clock concept . The genThe shorter drift distance of type 2 indicates it to be less attenuated than Sabin-1 and 3 viruses as far as transmission efficiency is concerned. Sabin-2 virus regains transmission efficiency fast if let loose among susceptible children. For Sabin-1 and 3 viruses, it takes longer; they regain neurovirulence early, but often without transmission efficiency. Globally cVDPV1 and cVDPV-3 emergence has been fewer and far between.Every dose of tOPV contains very small amounts of neurovirulent mutants ,11. TheyThese data had warned polio experts that Sabin-2 had to be very carefully managed as it could emerge as VDPV and cVDPV, if allowed to spread where and when population immunity was not high. High population immunity is the safety wall to block their emergence and spread. The original time frame for eradication was 1988 to 1999, when population immunity against type 2 was induced by both WPV-2 and tOPV . CoveragFrom the vaccinology viewpoint, that was the best time, indeed the only safe time, to withdraw Sabin-2, because of the wall of highest achievable population immunity ever. Such a moment occurs once in history, never to happen again. Sadly, knowledge, courage or wisdom did not prevail; tOPV was continued as business as usual, with the consequences that concern us today.There was another simpler reason why Sabin-2 had to be withdrawn. When a vaccine (like tOPV), with rare serious adverse reaction (namely VAPP) is used, benefit–risk balance had to be assessed. When WPV-2 was widely prevalent, the benefit of Sabin-2 was greater than its risk of VAPP. As WPV-2 was eradicated, the balance reversed: the risk from Sabin-2 vaccine remained, while benefit disappeared. Continuing Sabin-2 beyond its need, ignoring the negative benefit–risk balance, was against science and common sense.Polio caused by the Sabin virus (VAPP) or its variants is iatrogenic. The WHO had issued clear warning and guidance regarding VAPP in the 1980s ,17. CounEradication is precision public health—zero polio and zero poliovirus transmission. The end point was now obfuscated by not monitoring VAPP. Had it been monitored, Sabin-2 VAPP would have alerted countries of its risk without benefit. Should the GPEI have used OPV in the twenty-first century at all, when the eradication target was 2000? The predictable consequence of continuing OPV was VAPP, which went against the spirit of the resolution. When a country  or a WHO region  or a continent  was declared as polio-free, most people accepted it to be true without realizing that unknown numbers of VAPP would have been occurring.The emergence of VDPVs, cVDPVs, and polio outbreaks due to them, were not exactly predictable but their possibility was known, as we have mentioned in other sections. Obviously the GPEI took a risk and continued with widespread use of tOPV beyond 2000 and the rest is recent history.To withdraw type 2 vaccine virus from tOPV became a necessity. The GPEI waited 16 years to collect ‘evidence’ of polio due to VDPV-2 to be convinced. The probability of emergence of cVDPV lineages was to be expected due to gaps in type 2 immunity in children born since 1999. It was imperative that a wall of high population immunity be created before Sabin-2 withdrawal. The Mogilev experiment (described below) had given us sufficient warning about the silent persistence of Sabin-2 after its withdrawal, unless precaution were robust.From 2006 to 2015 many lineages of cVDPV were in circulation and all of them had to be interrupted. In 2016, polio was under eradication mode in a globalised world. There was only one way to build a robust wall of population immunity sufficient to interrupt all existing lineages and to pre-empt the emergence of any new lineages. It was to use both IPV and tOPV strategically. IPV had to be in 3 doses and given through EPI, achieving high coverage. Since six-antigen  vaccine including whole-cell pertussis and IPV was available, there was no need for additional injections of IPV. It could have been supplemented by multiple campaigns of tOPV, if the Mogilev experiment was clearly understood. After measuring and ensuring very high population immunity, Sabin-2 could be withdrawn, country by country. These were formidable tasks, but without them Sabin-2 withdrawal in 2016 was too risky.The GPEI was caught on the horns of a dilemma; cVDPV cases had occurred in 16 countries during 2009 to 2015, but proper preparations were labour-intensive and very expensive. The GPEI took a gamble—with one dose of IPV plus limited tOPV campaigns. Population immunity was not monitored. Sabin-2 was withdrawn from all countries irrespective of population immunity levels.When fresh cVDPV-2 lineages became visible after a lag time of one year, we knew that cutting corners was most unwise. Even when new lineages emerged in multiple locations, 3 doses of IPV were not introduced. No child anywhere has developed polio after taking 3 doses. Preventing polio in individual children and interruption of cVDPV-2 transmission were both important, but the GPEI neglected the former and tried its best to achieve the latter, so far in vain.In many countries, cVDPV is currently endemic. In the past four years 2018–2021) outbreaks of cVDPV-2 polio were recorded in 35 countries—25 in Africa and 10 in Eurasia—with polio cases totalling 2296 [–2021 outIn the Democratic Republic of Congo, cases have occurred every year; in Niger, Nigeria, Somalia and Pakistan over three years; in Afghanistan during 2020 and 2021. There were more cVDPV2 polio cases (513) in Afghanistan and Pakistan than of natural polio (354). When polio cases occurred due to importation of cVDPV-2 into Iran, Malaysia, Tajikistan, Ukraine and Yemen during these four years, there was hardly any discourse about it in international media.Families of children with cVDPV polio would not have realised that the tragedy was due to human error, not nature’s vagaries. No compensation was offered in spite of the root cause of human error. During these four years, cVDPV-1 polio cases were 90 in 8 countries and cVDPV-3 polio cases 7 in one country [“Those who cannot remember the past are condemned to repeat it”, said George Santayana. There was ample information in the twentieth century about the problem of the Sabin-2 virus mutating and becoming contagious and neurovirulent, if allowed to circulate. None of the successive GPEI leaders was experienced in polio epidemiology in low-income countries, nor well versed in the vaccinology of IPV and OPV. “Superficial knowledge is potentially more dangerous than ignorance. It gives a false sense of security encouraging an ignorant man to persevere in his efforts that can result in huge damage.” . What daOPV was introduced in the USA in 1961–1962 without assessing safety by clinical trial . VAPP waThe USSR had been using Sabin tOPV since 1959 in nationwide campaigns, followed by age-based routine immunisation. With abundant forethought, it planned an experiment in the Mogilev District in Byelorussia. After building up high population immunity through two nationwide OPV campaigns (covering 2 months to 10 years once and 7 years next), tOPV was not given in the entire district from March 1963 till March 1966 [The objective was to monitor the duration of immunity and duration of virus circulation in the absence of continued vaccination. Midway, in March 1965, 40 children in six nurseries were given one dose of tOPV. During May to October 1965, poliovirus type 2 was detected in 9 of 392 randomly selected children below 3 years—not from the six nursery cohort, but from others. One of them had facial palsy, which is a well-known form of polio .No type 1 or 2 virus was detected. Antibody studies showed wider prevalence of type 2 and limited prevalence of types 1 and 3 during the lull period. Vaccination with tOPV resumed in April 1966.This story was reviewed and data and viruses reanalysed in 2003 with clear advice not to reintroduce the type-2 vaccine once it was withdrawn . Yet, haSabin-1 or Sabin-3 vaccine could be withdrawn without (or with very little) risk of variants surviving in the community—but Sabin-2 could not be withdrawn at will, as it or its variants may already have been in transmission silently and may have ‘blown up in our faces’ as emerging VDPV and cVDPV. That is exactly what happened.We already said that it should have been withdrawn immediately after WPV-2 eradication. A wiser course was to begin planning for it in 1993 when the Egypt experience was fresh in memory. That was the ideal time to introduce IPV into EPI, with a view to wean the world off OPV by 1999 . Since tOne important lesson from the importations of VDPV-2 into London and New York, and its sustained spread locally over several weeks, is about the route of transmission. In London, the Sabin-related type 2 virus was in circulation for more than 6 months (February to July 2022) and in New York it has continued to circulate from March onwards ,26. The WPV and cVDPV, on the other hand, are contagious. Polio is a disease of infants and pre-school children with those infected having a median age below 15 months, typical of respiratory transmitted infectious diseases such as measles ,34,35,36This stark contrast is explainable as: OPV is adapted to intestinal infection, while WPV is adapted to nasopharyngeal infection as well as intestinal infection. It is during the period of nasopharyngeal virus shedding that WPV is transmitted to others. We are not aware of any research by the GPEI to explore if cVDPV-2 is pharyngeal-adapted. There is no observed or epidemiological evidence for faecal–oral transmission of WPVs ,38.The difference between the transmission efficiencies of OPV and cVDPV can be explained by the acquisition of transmissibility by the respiratory route, by deattenuation. Before Sabin promoted OPV, the general teaching was respiratory transmission, for which there is much epidemiological evidence ,38,39. HPolio was prevalent in all countries irrespective of standards of sanitation and hygiene. Sabin discovered that poliovirus infection occurred in the intestines, resulting in virus shedding in stools. He did not pursue the phenomena of pharyngeal infection, early upper respiratory symptoms of infected children, shedding via droplets/aerosol and the consequent contagiousness. The main argument Sabin put forth was the absence of WPV in the olfactory bulbs of children dying of severe polio to discredit the respiratory route of transmission .In communities highly endemic for WPVs and with very low median age of polio, Sabin OPV is not highly infectious—had it been otherwise, population immunity would have been near-100% by the time tOPV coverage was more than 50%. Infected children shed virus in stools for 4–6 weeks, but despite that vaccine viruses are not contagious. When one million infectious type 1 viruses were fed in tOPV, only 10% of infants in Northern India became infected . In spitThe epidemiology of polio in the pre-vaccine era was typical of respiratory-transmitted diseases. The textbook teaching was also that polio was due to respiratory transmitted infection ,43. All The importation of wild virus from Pakistan to Malawi and Mozambique in 2021 and the importation of VDPV-2 into London and New York and its sustained transmission are reminders that the assumption of the faecal–oral route is without any evidence. In both cases, it was probably adults who carried the virus to new territories; if so, reinfected adults were the likely transmission vectors. This will explain the paradoxical decline of WPV polio in Afghanistan and Pakistan in 2021 when all immunisations declined. In 2020 there were 149 cases but in 2021 only 6. When adults wore face masks against the coronavirus epidemic, WPV transmission declined drastically—however, this is only speculation.Obviously, Sabin OPV cannot be used in the polio-eradicated world, but IPV can be. If IPV is included in EPI, OPV can be withdrawn in any country that achieves high coverage—90% is reasonable as high coverage. The original World Health Assembly resolution stated: “…eradication efforts should be pursued in ways which strengthen the Expanded Programme on Immunisation…” . IPV as The first step in course correction of the GPEI immunisation tactics is the introduction of IPV, three doses, in all countries that are using OPV. The second step is withdrawal of OPV as stipulated above. Once OPV is removed, no new lineage of VDPV will emerge. As IPV will protect children against polio disease, we can wait for population immunity to interrupt cVDPV transmission. IPV offers better mucosal protection against respiratory transmission than OPV.Once OPV is globally withdrawn, any poliovirus, WPV, VDPV or Sabin-like, will be a signal of suboptimal population immunity that must be reinforced urgently. Continued environmental surveillance should then remain a crucial strategy to sustain a polio-free world. However, there will be no further need to classify each virus isolate by molecular analysis, thereby saving on expenditure.This paper is a revised version of a pre-print paper in Qeios .


We demonstrate the possibility of applying surface-enhanced Raman spectroscopy (SERS) combined with machine learning technology to detect and differentiate influenza type A and B viruses in a buffer environment. The SERS spectra of the influenza viruses do not possess specific peaks that allow for their straight classification and detection. Machine learning technologies  enabled the differentiation of samples containing influenza A and B viruses using SERS with an accuracy of 93% at a concentration of 200 μg/mL. The minimum detectable concentration of the virus in the sample using the proposed approach was ~0.05 μg/mL of protein (according to the Lowry protein assay), and the detection accuracy of a sample with this pathogen concentration was 84%. Outbreaks and epidemics of viral diseases in recent years have raised the active search for new methods for differential diagnostics and virus detection. Modern methods of virus detection, such as polymerase chain reactions (PCR)  and enzy3–109 times, and the subsequent detection of the pathogen in low concentrations (5 × 108 PFU/mL) [Surface-enhanced Raman spectroscopy (SERS) is considered a promising alternative for the differential detection of viral infections ,9,10,11. PFU/mL) . Identif PFU/mL) ,9,10,11. PFU/mL) ,13,14. T PFU/mL) . The lab PFU/mL) ,17.Many research efforts are underway to create a universal SERS substrate made from metal nanoparticles and nanorods which will allow us to determine the presence of viruses (including their type) in a sample ,20,21,22Spectral patterns of different influenza strains have similarities ,25 and tThe paper by Zhang Z. et al. described an experiment on modifying the structures of silver nanosubstrates by adding acetonitrile, bromine, and calcium ions; the minimum number of viruses for successful detection, in this case, was 100 particles per test . In theiThe work of Durmanov et al. describes the usage of a fabricated SERS substrate, which was composed of nanoporous mica with the addition of a thin silver layer by electron beam physical vapor deposition method . Four tyParia D. et al. reported on the creation of a label-free SERS platform with a metal-insulator-metal nanostructure . CombineSong C. et al. were able to demonstrate the possibility of using the portable Raman spectrometer to detect the influenza A virus . In theiThe support vector machine (SVM) method has been previously applied to the detection of varied biomarkers using spectroscopic data . A recenIn this paper, we propose implementing SERS and subsequent processing of the spectra to detect and differentiate the A and B influenza viruses in an STE buffer medium and investigate the limits of virus detection using this approach. Here, to the best of our knowledge, we, for the first time, demonstrate differentiating the A and B influenza viruses in an STE buffer based on SERS and SVM. We demonstrate successful detection and 93% accuracy in the differentiation of the viruses with low-cost commercial substrates, a simple STE buffer solution, and a standard machine learning algorithm that does not require time-consuming preprocessing steps of the spectral data. This makes our approach affordable and effective for use in real-life applications.Influenza A and influenza B viruses are typical agents causing acute respiratory infections. The antigenic structure of the hemagglutinin of influenza A and B viruses differs by ~70% , which mInfluenza A pdm09) and influenza B (B/Hong Kong/269/2017) viruses were grown in chick embryos; purified virus concentrate was then obtained by differential centrifugation of virus-containing allantoic fluid followed by ultracentrifugation reprecipitation. The Pierce BCA Protein Assay Kit  was used to measure the protein concentration in purified virus concentrates. The result of determining the concentration of protein, according to Lowry, in concentrated viral suspensions, was <200 µg/mL. The hemagglutination titer of the concentrate was 1:256. The specificity of the obtained viral concentrates was confirmed by the ELISA test using specific monoclonal antibodies to the hemagglutinating protein of influenza A and B viruses.The suspension of purified viral particles was placed in an STE buffer medium for further storage and use.The STE containing NaCl, Tris, and EDTA with pH = 8.0 was used as a buffer medium. The choice of this medium was justified by the absence of a fluorescence signal in the range required. The absorption spectrum of the STE buffer medium (in the visible and IR ranges) is shown in Commercial SERS substrates from SERSitive  were useThe measurements were carried out using a Horiba LabRam Raman spectrometer . The setup included a laser source , a spectrometer , a Mitutoyo Apo Plan 50x VIS lens, and a CCD camera . The design of the spectrometer is shown in The excitation radiation of the laser source was put through an aperture diaphragm serving as a spatial filter. The edge filter reflecting the excitation light and transmitting the Raman signal directs the excitation beam to the sample fixed at the object stage. The Raman scattering signal was then collected by the lens, passed through the edge filter, and directed to the spectrometer. The spectrally resolved signal was projected onto a CCD camera to obtain a Raman spectrum.It was experimentally found that the selected laser’s power and wavelength make possible the fluorescence signal reduction and allow for minimizing the probability of destroying the biological structures due to the nanoparticle heating.−1, which includes the main vibrational modes of organic compounds; the exposure time was 60 s. Several spectra were taken from five to six randomly selected spatial points on the substrate. As a result, 25–30 spectra were obtained for each sample, some of which are given in During the experiments, the first dark spectra and spectra of a pure substrate without a sample were obtained, with no Raman scattering peaks or artifacts visible during the process. Then, 1 µL volume samples of a clean STE buffer medium with influenza A and B viral particles , were applied to the SERS substrates surface. Further, the samples were dried at room temperature and placed in the spectrometer. SERS spectra were obtained in the range from 550 to 2000 cmThe Raman spectra of the influenza A and B containing samples are visually similar to the buffer medium spectra due to the strong signal from the latter. In this case, the differentiation of spectra and virus detection become complicated see , promptiTo determine the method sensitivity limits of the SERS spectra and classification using mathematical processing, the suspension with the influenza A virus was diluted with the STE buffer medium in various ratios: 1:10, 1:100, 1:1000, and 1:10,000. The initial protein concentration of the sample with the viral suspension was 500 µg/mL, according to the Lowry method . For thiThe experimental spectra obtained were combined into a table in which the columns corresponded to wavelengths and the rows corresponded to intensity values. The table that is used as a sample set had the dimensions of 57 × 1332 cells, where 29 lines corresponded to the spectra of the STE buffer medium and 28 lines corresponded to the spectra of the influenza A virus. For the mathematical processing, we used the package Scikit-learn v. 0.24.2 for Python v. 3.10.7 .The detection of the viral particles was carried out by binarily classifying the spectra of the pure buffer medium and the spectra of virus A in a buffer medium . For preliminary processing of the data, we applied standardization  and normalization  to the sample set. For classification, the support vector machine (SVM) method was used. The SVM is one of the most commonly used machine learning algorithms, the purpose of which is to solve the classification problem by constructing an optimal separating hyperplane .Samples that are closest to the hyperplane are the support vectors. The hyperplane is constructed so that the distance between it and the support vectors is maximized. This distance is called a margin. Accordingly, the remaining objects must have a distance to the hyperplane greater than the margin to perform the classification. This strict rule corresponds to the linear kernel.Cross-validation was carried out using StratifiedKFold .The average virus detection accuracy was 95.5% see . VisualiThe spectra of influenza A and B viruses were combined into a table and the total size was 90 × 1332 cells . An additional class-labeled column was introduced for classification purposes . Each class of spectra was represented by an equal number of samples (30 for each). Similarly, the methods of standardization  and normalization  of the samples were used for preprocessing. Here, we used the SVM method with the hinge loss kernel function for classification that introduces soft boundaries of a margin.Cross-validation was carried out using the StratifiedKFold method  set included per 27 (3) spectrum samples of the virus A, the virus B, and the pure buffer medium. Thus, a balanced division of the sample set into subsets with respect to the classes was obtained). In the full dataset, the mean cross-validation accuracy of classification was 93% . For difIn order to solve the binary classification problem of the pure buffer medium spectra from the influenza A virus spectra at different dilutions, the support vector machine with a linear classification kernel was used.For samples with different influenza A virus concentrations in the buffer medium , a data set was formed, which consisted of 10 measurements of the virus A spectra (for each concentration) and 10 measurements of the buffer spectra. Five subsets were formed for cross-validation . Thus, the test set for cross-validation had a dimension of 4 × 1332 (two spectra of the influenza A virus and two spectra of the pure buffer medium), and the training set was 16 × 1332 (eight spectra of the influenza A virus and eight spectra of the pure buffer medium).The model always accurately differentiated the virus and the buffer at high virus concentrations (the classification accuracy was 100%), but the classification accuracy decreased while lowering the concentration. The results of differentiation are presented in We determined the spectrum relative SD (RSD) to investigate the intensity signal deviation of the spectrum samples and their potential influence on the classification accuracy. We calculated the normalized intensity SD at each spectral wavenumber, normalized it by a mean value to obtain an RSD at each point, and then averaged it over the entire wavenumber range. The average RSD was 24% for 1:10, 42% for 1:100, 26% for 1:100, and 28% for 1:10,000. It should be noted that such relatively high values may be caused by using a label-free SERS substrate, and that no special techniques for ensuring uniformity of a sample distribution were undertaken. Despite these RSD values, no apparent influence of signal deviation was found during the classification process.4 times. At low concentrations, the method used was able to differentiate between virus-free and virus-containing samples with an accuracy of 84%. The results of the study will help to develop fast, cheap but reliable diagnostic methods for real life working with clinical samples.Through a combination with machine learning, photonics technology has flourished in biosensing by the implementation of extremely sophisticated methods to prepare biosamples, tailored substrates, and complicated preprocessing of spectral data for numerical analysis. In this work, we were focusing on design approaches to easy-to-use and affordable biosensing applications, and demonstrated the possibility of detecting and differentiating viral particles in a sample containing the STE buffer medium. The use of surface-enhanced Raman spectroscopy technology in combination with machine learning algorithms made it possible to differentiate samples with the influenza A virus, the influenza B virus, and without the virus, even for the limited data set sizes. The spectra were classified using the support vector machine. The accuracy of virus detection was 93%. To approach clinical studies, the concentration of the influenza A virus was reduced by 10