This study aimed to evaluate the efficacy and safety of trans-arterial chemoembolization (TACE) followed by stereotactic body radiation therapy (SBRT) in treating Barcelona Clinic Liver Cancer (BCLC) stage B hepatocellular carcinoma (HCC) not amenable to resection and radiofrequency ablation (RFA).From February 2012 to January 2017, a total of 57 BCLC stage B HCC patients who were unsuitable candidates for resection and RFA treated with TACE combined with CyberKnife SBRT were included in this retrospective study. Patients underwent TACE for a median of two times (1–5 times) before SBRT. SBRT prescription doses ranged from 30 Gy to 50 Gy in 3–5 fractions.The median follow-up time was 42 months. The objective response rate (CR + PR) was 85.9%, and the disease control rate (CR + PR + SD) was 96.5%. The local control (LC) rates were 91.1% and 84.3% at 1 and 2 years, respectively. The 1-, 2-, 3-year overall survival (OS) and the median survival time were 73.2%, 51.4%, 32.4% and 26.6 months, respectively. The 1-, 2-, and 3-year progression-free survival (PFS) were 34.2%, 21.6%, and 9%, respectively, with a median PFS time of 9.7 months. A subgroup analysis was conducted in 32 patients with AFP ≥ 200 ng/ml before TACE. OS was significantly prolonged in those with AFP that decreased by more than 75% than those with AFP that decreased by less than 75% (P = 0.018) after SBRT. The treatment was well tolerated with only one patient (1.8%) developed grade 3 gastrointestinal toxicity, and another patient developed non-classical RILD. In multivariate analysis, tumor length ≥ 10 cm and AFP ≥ 200 ng/ml were independent poor prognostic factors for OS.The combination of TACE and Cyberknife SBRT showed optimal efficacy with acceptable toxicity for BCLC stage B HCC. Primary liver cancer is the sixth most commonly diagnosed cancer, and its mortality rate ranks fourth around the world. According to the estimates of GLOBOCAN 2018 statistics produced by the International Agency for Research on Cancer of the World Health Organization, there are about 841,000 new cases and 782,000 deaths due to liver cancer annually. The ratio of death to new cases is as high as 0.9 , 4. All The National Comprehensive Cancer Network (NCCN) guidelines recommended that TACE combined with radiotherapy could improve the LC rate and prolong the survival time of patients with unresectable HCC, which was more effective than TACE and sorafenib , 6. HoweFrom February 2012 to January 2017, a total of 57 patients with BCLC stage B primary liver cancer who received TACE combined with CyberKnife SBRT treatment in Nanjing Jinling Hospital were included in this retrospective study. Inclusion criteria: 1. Patients were diagnosed as HCC by biopsy, and the imaging manifestations were nodular or lumpy; 2. BCLC stage B, Child–Pugh score A–B7 and ECOG 0–1; 3. Unsuitable for resection, liver transplantation, or local ablation therapies according to the comprehensive assessment of hepatobiliary surgery experts, oncologists, interventional experts, and radiologists; 4. Remaining healthy liver > 700 ml. Exclusion criteria: 1. Portal vein thrombus, lymph node involvement, and extrahepatic metastasis; 2. ECOG ≥ 2; 3. Poor liver function with Child–Pugh score of C; 4. Diffuse liver cancer or nonmeasurable lesion, tumor number ≥ 4; 5. Other life-threatening conditions, such as cardiac ischemia or cerebrovascular accident within the last 6 months.From February 2012 to January 2017, a total of 57 patients received the combined TACE and CyberKnife SBRT treatment as presented above. In our study, 44 (77.2%) patients were hepatitis B carriers. All patients had BCLC stage B disease. The median tumor size was 8.4 cm . None of the patients had previously received any other treatment, and no patient dropped out after TACE. The median number of TACE is 2 . The CyberKnife SBRT prescription doses ranged from 30 Gy to 50 Gy in 3–5 fractions. The median BED10 was 100 Gy . The median interval between TACE and SBRT was 37 days (23–69 days). Baseline patient and tumor characteristics are displayed in 2 cisplatinum, 20–40 mg THP, or 500–1500 mg fluorouracil glycosides was slowly injected through the catheter to the tumor site. The amount of the mixture emulsion should depend on the tumor size and arterial blood flow. Thereafter, gelatin sponge particle gelfoam embolization was conducted. Liver enhanced MRI and CT scans were performed 3 to 4 weeks after TACE to evaluate the lesion and short-term efficacy. TACE was repeated 1 to 5 times at intervals of 4 to 6 weeks. The median interval between the last cycle of TACE and CyberKnife SBRT was 37 days (23–69 days).TACE: Percutaneous puncture of the femoral artery with Seldinger technique was performed. The catheter was inserted into the hepatic artery or celiac axis under the guidance of DSA. Contrast agent was injected into the catheter to determine the location, size, number, and supply artery of the tumor. After the target lesion is determined, a catheter will be inserted to the feeding artery branch. A mixture of 5–20 ml of lipiodol and chemotherapy agents such as 30–40 mg/mCyberKnife SBRT: All patients were implanted with 3–6 gold fiducials (size of 6.0 mm × 0.8 mm) within or around the tumor using a CT-guided 19 G needle. A CT plain and enhanced scan was performed about 7 days after the implantation. At this time, edema and local hemorrhage subsided, and the gold fiducials were relatively stable and immobile.Patients were placed in a supine position and used a vacuum pad to fix the body. CT scanning was conducted, and the slice thickness was 1 mm. Hepatic scans ranged 15 cm above and below the lesions. The gross target volume (GTV) was defined as visible liver tumors at the arterial phase or at the delayed portal phase on the CT or MRI scan. The planning target volume (PTV) was defined as GTV plus a margin of 3 to 5 mm. After expansion, the area of the PTV should be adjusted according to the adjacent critical organs at risk. According to the tumor size, location, and critical organ constrains, patients were treated with prescription dose ranging from 30 Gy to 50 Gy for 3–5 times. Respiratory synchronization and gold standard tracking technology were adopted during the treatment. The prescribed isodose line should encompass >95% of PTV. Dose constraints for critical structures are shown in Enhanced upper abdominal CT and MRI were conducted 1 month after the completion of CyberKnife treatment, every 3 months in the first 2 years, and then every 6 months thereafter. According to the Modified Response Evaluation Criteria in Solid Tumors , the assSPSS 22.0 statistical software was used for data analysis. LC, PFS, and OS were calculated using the Kaplan–Meier method. Univariate analyses were used to investigate the relationship between all independent variables and OS. Any factors that were significant in univariate analyses were incorporated into multivariate analyses using the Cox proportional hazards model. P < 0.05 was considered statistically significant.After CyberKnife SBRT, liver MRI and/or abdominal CT was evaluated in all patients. CR occurred in 11 patients (19.3%), 38 (66.6%) patients achieved PR, 6 (10.5%) patients showed SD, and 2 (3.5%) cases developed PD. The objective response rate (ORR = CR + PR) was 85.9%. The disease control rate (DCR = CR + PR + SD) was 96.5%. The 1- and 2-year LC rates were 91.1% and 84.3%, respectively Figure 1As of the last follow-up date , the median follow-up time was 42 months . Four patients (7.0%) were lost to follow-up at the time of analysis, 1 patient (1.7%) underwent surgical resection, and 1 patient (1.7%) underwent liver transplantation, all of which were recorded as censored. The median OS was 26.6 months (95% CI 18.27–34.92), and the 1-, 2-, and 3-year OS were 73.2%, 51.4%, and 32.4%, respectively Figure 2The univariate analysis of OS identified five poor prognostic factors, including tumor diameter ≥ 10 cm, multiple nodules, AFP ≥ 200 ng/ml, BED10 < 100 Gy, and TACE times > 2. The significant factors of univariate analysis were enrolled in the multivariate analysis using the Cox proportional hazards regression model. The results showed that tumor diameter ≥ 10 cm and pretreatment AFP ≥ 200 ng/ml were associated with poorer OS Table 3.A subgroup analysis was performed on 32 patients with pretreatment AFP ≥ 200 ng/ml. Within 3 months after CyberKnife SBRT, the AFP of 18 patients (56.3%) decreased by more than 75%, and their OS was significantly longer than those whose AFP decreased by less than 75% (P = 0.018) Figure 4Patients mainly experienced grade 1 or 2 fatigue; nausea; vomiting; and hematological toxicity such as anemia, leukopenia, thrombocytopenia, hyperbilirubinemia, and AST elevation. Grade 3 or above adverse events included anemia (3.5%), leukopenia (5.3%), thrombocytopenia (7.1%), AST elevation (5.3%), and hyperbilirubinemia (7%). Three months after the treatment, no patient developed classic RILD, but one patient presented non-classic RILD. One patient developed gastric ulcer 5 months after treatment Table 4.The current guidelines formulated by the NCCN and American Association for the Study of Liver Diseases recommend TACE as the preferred treatment for inoperable HCC. According to the treatment algorithm of the BCLC, TACE is considered as the first-line treatment for intermediate-stage HCC patients who are not suitable for surgical resection or tumor ablation. However, TACE alone demonstrated a dismal CR rate of only 0%–4.8%, and patients cannot achieve satisfactory long-term survival, the 5-year cumulative survival rate of which is only 1%–8% . When thA meta-analysis  of 25 stIn recent years, an increasing number of studies have demonstrated that combination therapy benefits LC compared with monotherapy in the treatment of unresectable HCC , 24. In Clinical data have shown that TACE combined with SBRT in the treatment of unresectable HCC has better long-term survival than TACE or SBRT alone , 28. TifAlthough 30%–40% of primary liver cancer is negative for AFP , it is sWith regard to the side effects, TACE combined with CyberKnife SBRT was well tolerated, with the majority of adverse effects being grade 1 and 2. No patient exhibited classic RILD, but one patient had liver transaminase elevation without disease progression, which is defined as non-classic RILD. One patient developed gastric ulcer, whose target lesion was adjacent to the stomach. The maximum point dose reached 7 Gy*5fx, which might exceed the tolerance of the stomach. The toxic reaction could be gradually repaired after symptomatic treatment. The treatment-related adverse effects for most patients were tolerable, which may be due to the fact that 82.5% of the patients in this study had a Child–Pugh score of A. In addition, the interval between the last TACE and CyberKnife was more than 3 weeks, exceeding the time window of liver function repair, which was conducive to the recovery of liver function. Consistent with the opinion of most previous studies, TACE combined with SBRT was well tolerated in the treatment of HCC patients with Child–Pugh score of A/B7  found thThis study has several limitations. First, this is a retrospective cohort study from a single SBRT center, and the results might not be generalizable. Second, the number of enrolled patients is relatively small. Further randomized controlled studies are needed to address the limitations and determine the intended population, the most appropriate time for SBRT, the optimal number of TACE before SBRT, and the formulation of prescription dose and fractions in combination therapy. Recently, a randomized phase 3 trial (IMbrave150) showed that atezolizumab plus bevacizumab can prolong OS and PFS than sorafenib in patients with advanced unresectable BCLC B-C HCC . ImmunotIn conclusion, treatment with TACE plus CyberKnife SBRT was associated with optimal efficacy and acceptable toxicity in patients with unresectable BCLC B HCC.The original contributions presented in the study are included in the article/supplementary material. Further inquiries can be directed to the corresponding author.The studies involving human participants were reviewed and approved by Jinling Hospital review board. The patients/participants provided their written informed consent to participate in this study.CJ: project design, data collection, assembly, analysis, manuscript writing. SJ: data collection and assembly. HZ: data collection and assembly. AL: Investigation. XQ: data collection and assembly. XZ: project design. ZS: project conception and design, data interpretation. All authors contributed to the article and approved the submitted version.This work was supported by grants from: 1. The Nanjing Municipal Science and Technology Committee of Jiangsu Province, China (grant number: 201803050). 2. The Jiangsu Post-doctoral Research Funding Program, China (grant number: 2020Z305).The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


OBJECTIVES/GOALS: The goal of this study is to better understand the homicide victim population who were institutionalized within 30 days prior to their death. Improved knowledge of this population can potentially prevent these future homicides. METHODS/STUDY POPULATION: A retrospective analysis of the 36 states included in the 2003-2017 National Violent Death Reporting System was performed. Demographics of recently institutionalized homicide victims (RIHV) in the last 30 days were compared to homicide victims who were not recently institutionalized. Circumstances of the homicide, such as suspected gang involvement, were also compared. Parametric and non-parametric statistical analyses were performed. Significance was set at p<0.05. RESULTS/ANTICIPATED RESULTS: There were 81,229 homicides with 992 (1.2%) RIHV. The majority of RIHV were Black (49.6%) and older than victims who were not recently institutionalized . RIHV had a high school degree or higher in 54.8% of cases and the primary homicide weapon was a firearm in 67% of the deaths. They were more likely to be homeless (3.1% vs. 1.5%), have a mental health diagnosis (9.2% vs. 2.3%), abuse alcohol (6.1% vs. 2.2%), or abuse other substances (15.2% vs. 5.8%) [all p <0.001]. These victims were most commonly institutionalized in a correctional facility or a hospital compared to other facilities such as nursing homes. Homicide circumstances for RIHV were more likely to involve abuse/neglect , gang violence , or a hate crime (1.0% vs. 0.1%. p<0.001). DISCUSSION/SIGNIFICANCE OF IMPACT: Contact with an institution such as a hospital or prison provides high-risk patients the opportunity to potentially participate in violence intervention programs. These institutions should seek to identify and intervene on this population to reduce the risk of violent homicides.


However, the strength of the distance vs. VO2peak relationship is not strong, and there are no equations for estimating ventilatory threshold (VT), which is important for training prescription and prognosis. Since the 6MWT is often limited by walking mechanics, prediction equations that include simple additional predictors, such as the terminal rating of perceived exertion (RPE), hold the potential for improving the prediction of VO2max and VT. Therefore, this study was designed to develop equations for predicting VO2peak and VT from performance during the 6MWT, on the basis of walking performance and terminal RPE. Clinically stable patients in a cardiac rehabilitation program (N = 63) performed the 6MWT according to the American Thoracic Society guidelines. At the end of each walk, the subject provided their terminal RPE on a 6–20 Borg scale. Each patient also performed a maximal incremental treadmill test with respiratory gas exchange to measure VO2peak and VT. There was a good correlation between VO2peak and 6MWT distance (r = 0.80) which was improved by adding the terminal RPE in a multiple regression formula . The VT was also well correlated with walking performance, 6MWT distance (r = 0.80), and was improved by the addition of terminal RPE . The addition of terminal RPE to 6MWT distance improved the prediction of maximal METs and METs at VT, which may have practical applications for exercise prescription.Walking tests, such as the 6-min walk test (6MWT), are popular methods of estimating peak oxygen uptake (VO Specifically, the peak oxygen uptake  . Some equations ,22 also ce alone . 2peak. Following this line of thinking, Alamji et al. [However, because of the wide interindividual variability in the HR response and maximal HR, and of the profound effect of many medications on the HR response, it would be desirable to find a method of replacing the HR measure in these prediction equations. Eston et al.  have sugi et al.  have sug2peak and VT.Given that simple walking tests such as the 6MWT are much more assessable to the exercising community than laboratory-based tests, and that most training takes place at intensities just below the VT , this stn = 2), post myocardial infarction (n = 22), post revascularization surgery (n = 18), post percutaneous intervention (n = 14), stable heart failure (n = 7), and risk factors for cardiovascular disease (n = 25). The research protocol was approved by the University of Wisconsin-La Crosse Institutional Review Board for the Protection of Human Subjects . All subjects provided written informed consent before participation.The subjects for the study were 63 adult volunteers. All were participants in either a Phase II cardiac rehabilitation program or a community-based exercise program, designed for the primary and secondary prevention of cardiovascular disease. Diagnoses for patients were conventional: stable angina pectoris  and room air. Gas exchange was integrated every 30 s, and the highest 30 s VO2 was accepted as VO2peak. The VT was identified using both the v-slope and ventilatory equivalent methods [2prak and the VO2 at the VT (VO2@VT) was expressed as maximal Metabolic Equivalents (METs) (Max METs), and METs at the VT (METs@VT), as we felt that this was more clinically relevant and comparable to estimates of exercise capacity based on performance in standard exercise protocols [Each subject performed an incremental treadmill exercise test to fatigue or to clinical signs and symptoms  with conrotocols ,14,15,16rotocols . The disrotocols . InstrucThe relationships of 6MWT with Max METs and METs@VT were made using linear regression. Similarly, the relationships of the terminal RPE with Max METs and METs@VT were made using linear regression. Regression equations for predicting Max METs and METs@VT were constructed using multiple linear regression with a stepwise approach, with 6MWT distance entered first and terminal RPE entered second. The characteristics of the subjects are presented in r = 0.80) and METs@VT (r = 0.80), and a weak but statistically significant correlation between terminal RPE versus Max METs (r = 0.30) and METs@VT (r = 0.23).The bivariate relationships between 6MWT distance versus Max METs and METs@VT, and between terminal RPE vs. Max METs and METs@VT, are presented in 2 increased from 0.63 to 0.71, with a SEE of 1.3 METs and a standardized residual of 1.0 METs. The prediction equation was:When the 6MWT distance and terminal RPE were combined in a multiple regression equation to predict Max METs, the R2 increased from 0.64 to 0.69, with a SEE of 0.95 METs and a standardized residual of 0.7 METs. The prediction equation was:When the 6MWT distance and terminal RPE were combined in a multiple regression equation to predict METs@VT, the Rr = 0.87) and METs@VT (r = 0.85), there was a strong bivariate relationship for both. The residual scores (predicted-observed) revealed a small value for both Max METS (−0.27 ± 1.24 METs) and METs@VT (−0.14 ± 0.92 METs), with most of the outliers at a relatively higher exercise capacity  compares favorably with field tests using the 6MWT distance (R2 = 0.42) [2 = 0.78) [2 = 0.71) [2 = 0.80) [2 = 0.85) [2 = 0.71) [2max) is typically higher (R2 = 0.83–0.94), the SEE is also typically in the range of 1 MET [The goodness of fit of the equations for predicting Max METs from the 6MWT distance + terminal RPE (R = 0.78)  or R2 =  = 0.71) , the Coo = 0.80) , or with = 0.85)  or (R2 = = 0.71) . The SEEof 1 MET ,16,29,30The prediction of METs@VT has more typically been performed using percentages of the maximal power output , running−1 (2.5 mph or 4.1 kph). One would anticipate a RPE during training of ~13  [The workload required to achieve a particular value for the percentage of Max METS (%Max METS), percentage of HR (%HR) reserve, or METs@VT is probably lower than the workload eliciting these markers during exercise testing. Recent research from our laboratory  has sugg people)  and with people) . The tra people)  suggests−1 (4.5 mph or 7.2 kmh) which requires a VO2 of ~16 mL·kg−1·min−1 (~4.4 METs) [Another advantage of the 6MWT distance + terminal RPE approach is the ability to make outcome assessments in patients who are mechanically limited to a top walking speed. The 6MWT was originally created for patients with pulmonary disease or heart failure ,26, who .4 METs) . Since c2max might have huge significance, in particular, as most of the times it is hard to obtain cardiopulmonary exercise testing in a heart-failure population, and rough estimates based on 6MWT will be of paramount importance.Although the present study provides meaningful information, some limitations should be acknowledged. First, this study is a small cohort study. Therefore, future researched should consider a larger sample. Second, the present study did not include a large representation of the heart failure population  in which VOThe results of this study demonstrate that the simple approach of adding the terminal RPE to the 6MWT distance can improve the estimate of Max METs in patients in rehabilitation programs. Additionally, it can be used to make an estimate of the METs@VT, which is important prognostically ,11 as a


B19 (B19V) is a DNA virus, transmitted through respiratory secretions, hand-to-mouth-contact, blood transfusion and trans-placental transfer. It causes high mortality and morbidity in pregnant women, thus contributing to poor maternal and child health. B19V has been neglected due to dearth of epidemiological data. The aim of this study was to determine the sero-prevalence of Human Parvovirus B19 antibodies among pregnant women attending antenatal clinic at Federal Teaching Hospital Ido-Ekiti, Nigeria.Human Parvovirus This cross-sectional study enrolled pregnant women attending Federal Teaching Hospital Ido-Ekiti from January to May 2019 to obtain sero-epidemiological data. One hundred and twenty-two (122) consenting pregnant women were enrolled following institutional ethical approval. They were administered structured questionnaire and venous blood was collected in plain tubes for serum extraction. Sera samples were analyzed for IgG and IgM antibodies using the enzyme linked immunosorbent assay method. Percentages, median, chi-square and multivariate analysis were carried out using SPSS version 17.X2 =4.75). There was association between miscarriage, still birth, history of blood transfusion and prevalence of Human Parvovirus B19 (p<0.05).The prevalence of IgG was 44.3% (54/122), IgM 41.8% (51/122), and IgG/IgM 28.7% (35/122) leaving 55.7% (68/122) of the population uninfected. The median age of participants was 22 (Interquartile range 18-25) years among which 36-45years had the highest prevalence which was not statistically significant (p=0.09 There is a high Prevalence of B19V among pregnant women attending antenatal clinic in this study. This underscores the need for testing and immunization of pregnant women against B19V. B19V has been linked as a cause of hydrops fetalis, intrauterine fetal death, aplastic crisis, spontaneous abortion, acute symmetric polyarthropathy and erythema infectiosum (Fifth disease) . Its discovery was first documented in 1975 .Human Parvovirus B19 (B19V) is a 5.5kb single stranded, non-enveloped DNA virus, transmitted through respiratory secretions, hand to mouth contact, blood transfusion and trans-placental transfer  and fetal death . Fetuses at first and second trimester are most susceptible because erythroid precursor development is predominant at these stages . Of the three genotypes described worldwide, type 3 is reported to be the most commonly occurring in the North and West Africa . B19V has specific tropism for blood group Antigen P (globosid) which is expressed on the surface of erythrocyte precursor cells, megakaryocytes, endothelia cells, placenta cells and cells at fetal myocardium . Human Parvovirus B19 infects erythrocyte progenitor cell stimulating apoptosis of the infected cells. Infections with this virus are usually asymptomatic in most patients but acute infection in patients with a high erythrocyte turnover may lead to a life-threatening aplastic crisis. For instance, acute congenital B19V among pregnant women in developing countries is not well documented, few available documents show a sero-prevalence of 3.3% for B19V IgM in South Africa , 32.8% in Tanzania  and 13.2% in Central Nigeria . A prevalence of 24.9% for B19V IgG has been documented in South Africa, 55.0% in Tanzania , 27.5% in Central Nigeria  and 20% in South-West Nigeria .Though the prevalence of B19V despite its immense impact on child and maternal mortality was not enlisted . This is possibly due to paucity of epidemiological data on impact of B19V on pregnant women in Nigeria. The aim of this study was to determine the prevalence and impact of Parvovirus B19V in pregnant women.Nigeria has a high maternal and child mortality rate which has become the main focus of the government in health care delivery in recent years. A number of causes for these mortalities have been listed; unfortunately, ..\Documents\Find coordinates.docx. Laboratory analysis was carried out in the Medical Laboratories of the Department of Medical Laboratory Science, Afe Babalola University, Ado-Ekiti (ABUAD), Ekiti State. Ado-Ekiti is a city in Southwestern Nigeria and lies on latitude 7o 35 and 7o38 north of the equator and longitude 5o10 and 5o15 east of the Greenwich Meridian . Afe Babalola University is a private institution with its campus located at Km. 8.5, Afe Babalola way, opposite Federal Polytechnic, Ado-Ekiti.This cross-sectional study was conducted between January and May 2019 at the Federal Teaching Hospital Ido-Ekiti (FETHI), Ekiti State, among pregnant women attending antenatal clinic. Federal Teaching Hospital Ido-Ekiti is a referral hospital for Ado-Ekiti and some towns in neighboring states such as Kogi, Kwara and Osun states. It is located in Ido-Osi Local Government Area of Ekiti State Nigeria, latitude 7.843093, longitude 5.182314 The study population included one hundred and twenty-two pregnant women attending antenatal clinic at the Federal Teaching Hospital Ido-Ekiti. Subjects who consented to take part in the study were randomly selected and administered a structured questionnaire to collect social demographic and some health history data.Ethical approval was obtained in accordance with the Helsinki declaration from the Ethics and Research Committee, Federal Teaching Hospital, Ido-Ekiti, Ekiti State (Protocol No: ERC/2019/03/13/199B). The study participants were informed about the purpose of the study and written consent was obtained from each participant before sample collection.th week were excluded.Pregnant women at exactly their 36Those with gestational diabetes and high blood pressure were excluded.The minimum sample size for this study was calculated using Yamane Taro’s formula (1967) for finite population .Where,n= minimum sample sizeN= average number of pregnant women attending FTH IDO-EKITI in six monthse= level of precisionN= 198e= 0.05One hundred and twenty-two (122) 5ml venous blood samples were obtained from consenting participants after the administration of a structured questionnaire into plain tubes. Samples were centrifuged at 1000 rpm (revolution per minute) for 10 minutes. The Sera were harvested using Eppendorf tubes and stored at -20°C .The sera samples were used to detect parvovirus B19V IgG and IgM antibodies using ELISA kit : according to manufacturer’s instructions.oC for 60 minutes . After incubation, the microplate was washed five times using wash buffer. 100 µl of Horseradish peroxidase enzyme (HRP) conjugate was added to each well except the blank; the microplate was covered with a sealing paper and also incubated in a microplate incubator at 37oC for 15 minutes. After incubation, the microplate was washed five times with the diluted wash buffer in an automatic plate washer  . 50 µl of substrate solution A and B were added to each well respectively and were mixed; the plate was covered and incubated at 37oC for 15 minutes. 50 µl of stop solution was added to each well and mixed. The absorbance was read in an ELISA reader machine  at a wavelength of 450 nm .Briefly, the test procedures were as follows: The reagents provided were allowed to attain room temperature for 15 minutes before use. The 20X wash buffer was diluted with distilled water using a ratio of 1:20 before use. The micro-titer plate template was set up with 1 well as blank, 2 wells as negative control and 2 wells as the positive control. 10µl of sera sample and 40µl sample diluent were dispensed into the respective wells except for the blank well, negative control well and positive control well. 50 µl of the negative and positive controls were dispensed into their wells respectively. The content was mixed by vibrating the plate gently. The microplate was covered with a sealing paper and incubated in a microplate incubator  at 37If mean negative control O.D < 0.1 and the mean positive O.D > 0.8, the test is valid.Cut off = the mean O.D value of the negative control × 2.1Positive results: Sample O.D > Cut-off O.DNegative result: Sample O.D < Cut-off O.DB19V in pregnant women.Collected data were analyzed using statistical package for social science (SPSS) version 17. Categorical variables were summarized as proportions while continuous variables were summarized as median with interquartile range. A multivariate regression was also conducted to determine predictors of seropositivity of B19V IgG antibodies while 51/122 (41.8%) were seropositive for B19V IgM antibodies leaving 68/122 (55.7%) as the susceptible population  were seropositive for B19V IgM only .Participants had a median age of 22(IQR 18-25) years. Out of the 122 subjects, 65 (53.3%) were age group 26-35 years. 11% and 55% of the subjects were in their first and second trimester respectively while the remaining 34% were in the third trimester. Twelve percent (12%) of them had a history of still birth or miscarriage. A total of 54/122 (44.3%) of subject were seropositive for pulation . Thirty-IgM only . PrevaleB19V. There was no association between the stage of pregnancy and the distribution of B19V (p = 0.90 for IgG and 0.32 for IgM). There was an association between history of blood transfusion, complications and the seroprevalence of B19V as those who had history of blood transfusion (OR 2.1 95% CI 1.7 - 2.3) p = 0.05, miscarriages (OR 2.5 95% CI 1.7-3.3) and still births (OR 2.8 95% CI 1.7–3.1), p = 0.04 for IgG and (OR 2.1 95% CI 1.1–2.7) and (OR 1.7 95% CI 1.1–1.9) respectively, p = 0.05 for of IgM (B19V (For the two vulnerable stages of pregnancy (first and second trimester), 4.1% (5/122) and 23% (28/122) respectively were seropositive for IgG while 3.3% (4/122) and 26.2% (32/122) respectively were seropositive for IgM antibodies leaving 44% of the population susceptible to r of IgM . Parity,gM . This present study was in like manner conducted between January and May, 2019. It is also possible that the year 2019 when the study was conducted coincides with the epidemic year of B19V. This argument is backed up by the report of Ender and colleagues (2007) which suggested that the epidemic cycle of B19V appears to be around 4 years with 1-2 epidemic years followed by 2 or 3 years of less frequent rates of infection. w. However, the B19V IgG/IgM co-prevalence which Mirambo et al., referred to as true indicator of incidence was 28.7% in this present study. The prevalence reported is significant because 44.3% of the population do not have antibodies against B19V as such if they get infected during a first or second trimester of pregnancy, they may develop complications.The findings of this present study revealed 44.3% B19V IgG prevalence and 41.8% B19V IgM prevalence among the study population. The prevalence as indicated by B19V IgG is similar to the 41.5% reported in Northern Nigeria , 52.5% reported in North Central Nigeria  and 55% reported in Tanzania  but higher than 20%  reported in Southwest Nigeria. The increased prevalence from Southwest Nigeria could be related to time lag since the research was conducted about 10 years ago. This suggests that the prevalence of B19V has doubled over the last ten years. The incidence of the disease as indicated by B19V IgM was very high similar to 39.9% reported in India  but higher than 13% reported in Central Nigeria  and 9.5% in Southwest Nigeria. This disparity may not be unconnected to seasonal peaks variation and the evolving epidemiology of the virus. It has been reported that the peak period of B19V is usually late winter and spring with 75% of IgM-positive cases recorded between January and June , the highest prevalence was recorded among the age group >36 years but contrary to their report, prevalence of IgM was also highest in the lower age group in this study. The reason for this may not be unconnected to cultural and behavioral diversity in different locations of the country. Similar to the report by Abiodun et al. (2013) there was no association between age, Incidence (IgM) and prevalence of B19V (IgG) recorded in this study.Similar to the findings in North central Nigeria , we found no association between parity and prevalence of B19V. There was an association between history of blood transfusion and the prevalence of both IgG and IgM (P = 0.04 and 0.05 respectively). Based on the association recorded, IgG had an odds ratio of 2.1 (95%CI 1.7-2.3) which indicates that those who had B19V had a 2.1 times likelihood of haven been exposed to blood transfusion. In like manner, IgM had an odds ratio of 2.5 (95%CI 1.8-2.9) also implying 2.5% likelihood of haven been exposed to blood transfusion. This finding corresponds with previous reports  that documented an association between Blood transfusion and prevalence.Seventy-one percent (36/51) of subjects with recent infections as indicated by the presence of IgM where still within the first or second trimester; a high-risk stage for B19V screening be included as a criterion for blood donation and transfusion. An association was recorded between history of pregnancy related complications and prevalence of both IgG and IgM (P= 0.04 and 0.05 respectively). These complications include Miscarriages and still births. Their odds ratio suggests that women who had suffered miscarriage and still birth had greater than two times likelihood of have been exposed to B19V infection. This finding deferred from the report of Okojokwu et al. (2018), who reported no association between miscarriage, still birth and prevalence of B19V. This is probably due to sociocultural variations between the two study locations. There was no association recorded between parity, number of pregnancies, occupation, marital status and prevalence of B19V IgG and IgM. The high incidence rate recorded within a population with a high percentage of susceptible individuals is indicative of an epidemiologic risky situation requiring a strategic intervention plan.It is hereby recommended that In the present study, we recorded a 44.3% B19V IgG and 41.8% B19V IgM prevalence rate leaving 44.3% of the population with the potential of getting infected with the virus. The findings point to a possibility of association between history of blood transfusion, history of complication and both prevalence and incidence of B19V. This is indicative of an urgent need for a B19V intervention plan which should include routine laboratory testing and immunization against B19V among pregnant women or intending mothers.B19V- Human Parvovirus B19IQR- Interquartile rangeOR- Odd RatioNIHF- Non-immune hydrops fetalisCI- Confidence interval


It can affect both fertility and the course of pregnancy. The aim of the study was to investigate the influence of pregnancy as a factor of recurrence or progression, on the prognosis of thyroid carcinoma.we conducted a retrospective cohort study of 117 young female patients followed up for papillary thyroid carcinoma (PTC) at the Department of Endocrinology, Diabetology and Metabolic Diseases of Ibn Rochd University Hospital of Casablanca, from January 2010 to December 2018, divided into 2 groups: group 1 composed of pregnant patients (n=42) and group 2 being the control group (n=75). Statistical analysis was made using SPSS software version 22.0.0.average age of pregnant patients was 35 ± 6.5 years old. Mean duration between first pregnancy and treatment completion was 4.4 ± 3.1 years. Over an average treatment duration of 14.4 months in postpartum, 30 patients were in remission (thyroglobulin (Tg) <1μg/l, negative thyroglobulin antibody and no morphological abnormality), while 12 had persistent symptoms . Thyroid stimulating hormone (TSH) during pregnancy was on average 0.83 mIU/l. Cancer progression was correlated with persistence of thyroid cancer to treatment before pregnancy (p = 0.01), pre-existing distant or locoregional metastases (p = 0.02) and delayed administration of radio-iodine therapy (p = 0.01). Interval between diagnosis and pregnancy, TSH level during pregnancy or the pre-conception thyroglobulin level did not have a statistically significant impact. Pregnancy was not associated with progression or recurrence of thyroid cancer .this study shows that pregnancy has no impact on recurrence or progression of thyroid cancer in patients declared in remission prior to conception. The incidence of thyroid cancer is increasing worldwide . PapillaWe conducted a retrospective cohort study from January 2010 to December 2018, on the patients followed up for papillary thyroid carcinoma. The recruitment site was the Endocrinology Service - CHU Ibn Rochd Diabetology of Casablanca.Inclusion criteria: our study included 117 patients selected according to the following criteria: age <45 years, total thyroidectomy; radioiodine (I-131) therapy indication according to clinical stage; follow-up duration over one year. Our patients were divided into 2 groups: Group 1 (n = 42) experienced pregnancy during postoperative follow-up, group 2 (n = 75) patients who have not been pregnant during follow-up. Thus, patients in group 1 with PTC were followed up for thyroid carcinoma since the pregnancy diagnosis until delivery, with regular postpartum monitoring.Variables studied: parameters studied were: interval between completion of treatment and pregnancy, duration of follow-up, course of pregnancy, obstetrical prognosis, thyroid status in pre/per/postpartum, and postpartum evolution of carcinoma. Each patient was classified according to the World Health Organization classification [th edition of the TNM classification (American Joint Committee on Cancer (AJCC)) [fication , organiz (AJCC))  with a r (AJCC)) . To deteRemission: suppressed Tg below 1 ng/ml, or undetectable ultrasensitive Tg (< 0.2 ng/ml), with negative thyroglobulin antibodies (TgAb) and without morphological abnormality on ultrasound.Recurrence/progression of the disease were defined by: detectable Tg levels (>2 ng/ml) or persistent TgAb .Data collection and analysis: data was collected from medical files, then transferred into an Excel sheet. We used SPSS software, version 22.0 to analyse data. For the descriptive part, the standard deviations, means, and percentages, were calculated to summarize the distributions of qualitative variables in pregnant and non-pregnant women. Pearson Chi 2 test was used to evaluate the association between clinical and obstetrical factors with the progression of thyroid carcinoma; the significance threshold (p) was set at < 0.05. Factors identified as significant by univariate analysis were further investigated using logistic regression.Our study included 117 patients with PTC screened from January 2010 to December 2018. The average age in Group 1 (G1) was 35.0 ± 6.5 years, which was statistically higher than the control group (G2), which was 31.2 ± 5.7 years (p = 0.02). All patients underwent thyroidectomy with a levothyroxine substitution after surgery. In only six women (14.2%), TSH levels was less than 1 mIU/L during pregnancy. In 36 women (85.7%), at least one TSH measurement was above 3 mIU/L during pregnancy. Mean TSH level during pregnancy was 0.83 ± 2.15 mIU/L. However, no correlation was found between TSH levels and progression of the disease or thyroglobulin during pregnancy. No significant differences were noted in tumor size nor in extrathyroidal extension (Tumor-node-metastasis (TNM) classification). No deaths were noted. The average time between pregnancy and completion of treatment was 4.4 ± 3.1 years (range 1 to 8) .Course of pregnancy: the pregnancy was uneventful in all cases except one, which ended in a miscarriage. The most common delivery mode was cesarean section in 59% of cases. The average birth weight was 2865 ± 475 g. Average follow-up duration after the first delivery was 14.45 ± 2.4 months.Evolution: in postpartum, 30 patients were in remission, while 12 patients had persistent cancers, without significant difference between the two groups (p = 1.25) In the 42 women, the average Tg before pregnancy was about 1.75 ng/ml (range: 0 to 12.4 ng/ml) and the average rate after delivery was 1.98ng/ml (range: 0 to 18.5 ng/ml). The difference was not statistically significant (p = 1.00). Persistent cancer was detected in 12 of the 42 pregnant women (28%), one of them showed disease progression after delivery (2.4%) (p = 1.00). Distant metastases were detected in 3 of 42 pregnant patients (7%) and 18 of 75 non-pregnant women (24%) with a significant difference observed between the 2 groups (p = 0.01). Multivariate logistic analysis was performed on the general population (including two groups: group 1 versus the control group). Relative risks (RR) were adjusted for age at diagnosis of disease. Univariate analysis (analysis  indicateanalysis .et al. [Differentiated thyroid cancer is more common in women. It is the second most diagnosed cancer during pregnancy and postpartum period, with a prevalence of 14 per 100,000 live births . Severalet al.  have comet al. [et al. [et al. [A retrospective study by Budak et al.  was perf [et al.  showed,  [et al.  tested 6The interval between diagnosis and pregnancy, the TSH levels during pregnancy or thyroglobulin level before conception had no statistically significant impact on the progression of thyroid carcinoma. Anxiety in cancer patients who wish to become pregnant is very high . In our Pregnancy has no significant effect on the recurrence of thyroid carcinoma in patients exhibiting no persistence of the disease before conception. Only a delay in treatment, in particular a delay in radioiodine therapy, could affect the prognosis in patients not declared remitted before pregnancy. However, regular monitoring of patients with thyroid cancer in pregnancy is still important.Thyroid carcinoma is more frequent in women of reproductive age;It can affect both fertility and the course of pregnancy.The study established that pregnancy has no effect on the recurrence of thyroid carcinoma in patients exhibiting no persistence of the disease before conception.


Dementia is a complex syndrome with various presentations depending on the underlying pathologies. Low emission of transcranial near-infrared (tNIR) light can reach human brain parenchyma and be beneficial to a number of neurological and neurodegenerative disorders. We hereby examined the safety and potential therapeutic benefits of tNIR light stimulations in the treatment of dementia. Patients of mild to moderate dementia were randomized into active and sham treatment groups at 2:1 ratio. Active treatment consisted of low power tNIR light stimulations with an active photobiomodulation for 6 min twice daily during 8 consequent weeks. Sham treatment consisted of same treatment routine with a sham device. Neuropsychological battery was obtained before and after treatment. Analysis of variance (ANOVA) was used to analyze outcomes. Sixty subjects were enrolled. Fifty-seven subjects completed the study and had not reported health or adverse side effects during or after the treatment. Three subjects dropped out from trial for health issues unrelated to use of tNIR light treatment. Treatment with active device resulted in improvements of cognitive functions and changes were: an average increase of MMSE by 4.8 points; Logical Memory Tests I and II by ~3.0 points; Trail Making Tests A and B by ~24%; Boston Naming Test by ~9%; improvement of both Auditory Verbal Learning Tests in all subtest categories and overall time of performance. Many patients reported improved sleep after ~7 days of treatment. Caregivers noted that patients had less anxiety, improved mood, energy, and positive daily routine after ~14-21 days of treatment. The tNIR light treatments demonstrated safety and positive cognitive improvements in patients with dementia. Developed treatment protocol can be conveniently used at home. This study suggests that additional dementia treatment trials are warranted with a focus on mitigating caregivers’ burden with tNIR light treatment of dementia patients. This number is projected to reach 13.8 million in the year of 2050 [Dementia is a complex syndrome with different clinical presentations. Although the course of disease and its clinical progression vary largely contingent on the underlying pathologies, the development of brain dysfunction that typically progresses to diffuse deficits is characteristic . The mosLow power transcranial near - infrared (tNIR) light emitting diodes (LED) illuminate light which is outside of visible spectrum of human eyes but can efficiently penetrate skin and skull to reach brain parenchyma -5. RecenIn this study we examined the safety and potential therapeutic benefits of tNIR light stimulation in the treatment of patients with dementia.This placebo controlled, randomized, double-blinded study was approved by institutional IRB, performed under protection embodied in the Basic Principles of the Declaration of Helsinki, and conducted at Baylor Scott & White Health (BSWH) Medical Center in Temple, TX. Sixty patients diagnosed with early and moderate dementia were enrolled and randomized to the active arm or control arm at a 2:1 ratio. Subjects, family caregivers, and investigators were masked. Inclusion criteria: 1) patients of all sex, age 50-85-year-old; 2) diagnosed with early- to mid-stage dementia or dementia-related symptoms; 3) generally healthy as indicated by recent physical examination within the last 6 months. Exclusion criteria: 1) diagnosed with actively growing, or a history of recurrent intracranial neoplasms; 2) history of epilepsy; 3) history of acute ischemic/hemorrhagic stroke.2, ~650cm2 per treatment area. Treatment protocol was two 6 minutes sessions daily for 8 consequent weeks at home with either active or sham device self-administered by patient/family caregiver.Both active and sham light treatment helmet devices had 12 cranial modules with 70 LEDs/module and 2 foldable eye modules with 14 LEDs/module. Sham helmet devices designed identical to active devices but did not emit NIR light. Active helmet devices emitted low power NIR light with wavelength of 1060-1080nm and 15,000mW, irradiance or power density= 23.1mW/cmNeuropsychological assessments of behavior, mood and cognitive performance were conducted at beginning (before the treatment) and at the end (within one day after the last treatment session) of study. Alzheimer’s Disease Neuroimaging Initiative (ADNI) neuropsychological battery was implemented to assess cognitive function of participants. ADNI battery included: 1) mini-mental state exam (MMSE) - a cognitive screener that briefly measures orientation, word recall, attention, working memory, copying skills, and abilities; 2) ADAS-Cog ; 3) clock drawing test (CDT) - subjects asked to draw a clock to a requested time; 4) logical memory (immediate) - subjects asked to recall a story immediately after it is been read; 5) auditory verbal learning test- immediate (A.V.L.T.-1) - subjects asked to memorize and recall immediately list of words presented verbally in several trials; 6) digit span forward and backward (DSF and DSB) - subjects asked to recall given different sequences of numbers in same order (forward span) or reverse (backward span).; 7) category fluency test - subjects asked to name words in given category within one minute; 8) trail making tests A and B - subjects asked to connect circles in numerical and mixed  order, respectively in limited time; 9) WAIS-R digit symbol substitution test - a cognitive test to assess visual motor coordination, motor persistence, attention and response speed; 10) Boston naming test - subjects asked to name 30 objects/items printed in the book; 11) logical memory (delayed) - subjects asked to recall a story after 30 min delay; 12) auditory verbal learning test- delayed (A.V.L.T.-2) - subjects asked to recall list of words presented verbally 30 min prior the recording in several trials. Daily subjective responses record was documented by caregivers. Sleep associated findings were collected from caregivers’ daily logs and feedback notes from patients/caregivers during assessment visits. All assessments/evaluations were performed by the same examiner using same procedures across the study.p < .05.Analysis of variance (one-way ANOVA) was used to assess mean differences between testing occasions and analyze outcomes inside of each group. Clock drawing and clock copying tests were scored using Shulman method. Analyzed data were considered to be statistically significant when Total of 60 subjects were enrolled into the study. Three subjects withdrew from the study due to health issues unrelated to the use of tNIR light therapy. Fifty-seven enrolled patients successfully completed the 8 weeks study course. Mean age of overall study population was 74.2 ± 7.7 years old with 60% of male and 40% of female distribution. Mean age of population treated with active device was 72.4 ± 8.2 years old with 59% of male and 41% of female distribution. Mean age of population treated with sham device was 77.8 ± 5.2 years old with 53% of male and 47% of female. Patients had no health or adverse effects reported during or after completion of study associated with use of tNIR light stimulation. Notably, both patients and family caregivers from the active arm shared positive feedback of noticeable changes in cognition and improved daily routine activities.In the active arm, some patients appreciated longer and more peaceful night sleeps. Duration of sleep increased by 1 hour in average after 8 ± 2 days of the treatment. Recurring nightmares ceased in some patients receiving tNIR light therapy. Patients reported being more energetic, physically and mentally engaged in daily living. Caregivers noted that patients had less anxiety, improved mood, energy, and positive daily routine after approximately 14-21 days of treatment, which was not noted by caregivers in the placebo arm.When results of active tNIR light treatments were compared with placebo effects in the end of trial, several improvements were noted as follows:p < .001) at the end of the treatment, which was 4.8 points improvement (21.0% increase) over the course of treatment. In the control arm, the average MMSE score changed from 23.2 ± 1.6 at the beginning of treatment to 24.6 ± 2.5 (p = .066) at the end of the treatment, which was 1.4 points improvement (6.2% increase) over the course of treatment . In the active arm, the average MMSE score improved from 22.8 ± 2.6 at the beginning of treatment to 27.6 ± 2.8 (reatment .p = .08) at the end of the treatment. This was 0.7 points improvement (20.5% increase) over the course of treatment. Patients with sham treatment did not demonstrate significant changes in CDT with average scores shifting from 3.75 ± 1.39 to 3.81 ± 1.38 (p = .89), which is a mere 1.7% change in mean CDT score over the course of sham treatment (Clock Drawing Test (CDT). Positive trends of improved CDT scores were recorded in patients received active tNIR light treatment with mean value going up from 3.5 ± 1.5 at the beginning of treatment to 4.2 ± 1.2  at the end of the treatment resulted in 8.7% increase over the course of treatment. Subjects treated with sham devices demonstrated decrease of CCT performance from an average of 4.8 ± 0.8 at the beginning to 4.7 ± 1.0 (p = .84) at the end of the treatment (Clock Copying Test (CCT). Subjects with active tNIR light treatment showed trend of mean CCT score value improvement from 4.2 ± 1.3 at the beginning of treatment to 4.6 ± 0.9 (reatment .p = .03) at the end of the treatment and resulted in 3.3 points increase over the course of treatment. In comparison, placebo effect on execution of logical memory- I test demonstrated decrease in performance of story recall with average score shifting from 7.4 ± 5.0 at the beginning to 6.0 ± 4.5 (p = .39) at the end of the treatment. This resulted in 1.4 points decrease over the course of treatment . Total story passage recall/learning trial in patients with active tNIR light treatment demonstrated improved performance. Average story recall scores increased significantly from 8.5 ± 5.9 at the beginning of treatment to 11.8 ± 7.0 (reatment .p = .06) at the end of the treatment. Sham treated group demonstrated decrease in performance similar to LMT-I test and resulted in test score decrease by 1.6 points from 4.3 ± 4.6 to 2.7 ± 4.0 (p = .29) over the course of treatment . Active device treated group also demonstrated improved performance by increase of average score by 3.0 points from 6.5 ± 5.9 at the beginning of treatment to 9.5 ± 7.8 (reatment .p = .03) at the end of the treatment which was 16.0 seconds (23.5%) faster than the average performance before the treatment. This finding is in stark contrast to sham treated patients’ performance which demonstrated a broad range of responses from no change to slower performance. Average time to completion of sham treated dyad changed from 68.4 ± 35.5 seconds at the beginning of treatment to 70.6 ± 41.4 seconds (p = .87) at the end of treatment. This resulted in average of 2.2 seconds (6%) slower performance for the task when compared to results before sham treatment  at the end of treatment, which is 40.4 sec (23.7%) faster performance compared to the speed of execution prior to the treatment. On other hand, sham treated patients, once again, demonstrated trend of slower execution after completion of treatment course. In this group, an average time of completion changed from 167.9 ± 80.8 seconds to 176.6 ± 88.0 seconds (p = .80) resulted in slower performance by 8.7 sec (5.2%) when compared with time of performance before the treatment  at the end of treatment, which is 2.2 points increase (8.8%) compared to levels before treatment. Some responses to the treatment resulted in up to 35.3% of score improvement. These results were noted in three cases in patients diagnosed with moderate dementia. Sham treated patients demonstrated insignificant BNT average score change from 23.4 ± 4.6 to 24.3 ± 4.9 (p = .58), which is 0.9 points shift (4.0%) compared to the level before treatment.Boston Naming Test (BNT). Patients treated with active device had significantly better average performance on BNT compared to sham treated group. Average score in this group improved from 24.4 ± 4.4 at the beginning to 26.6 ± 3.8 (p = .15) at the end of treatment, resulted in 0.7 points (10.3%) increase. In the active group, DSB test average score shifted from 4.8 ± 1.7 to 5.5 ± 2.1 (p = .17), a 0.6 point (12.4%) increase over the course of treatment. In the control group, neither DSF nor DSB had significant change in test performance compared to levels prior the treatment. Average test scores shifted from 6.4 ± 1.9 to 6.4 ± 2.0 (p = .92) and from 5.0 ± 1.4 to 5.0 ± 1.2 (p = 1.0) for DSF and DSB tests, respectively.Digit Span Forward (DSF) and Digit Span Backward (DSB) Tests. Active device treatment resulted in positive change of DSF test average score from 6.9 ± 2.2 at the beginning to 7.6 ± 2.2 (p = .19), which is 3.8 (12.6%) points higher than before the treatment. Placebo effect did not trigger significant improvements of performance on this test and resulted in change of average score from 29.25 ± 13.9 to 29.8 ± 14.8 (p = .91), which is a mere 0.6 (1.9%) points change over the course of treatment.WAIS-R Digit Symbol Substitution Test. Treatment with active device resulted in trends of improvement of performance from 29.9 ± 11.4 to 33.7 ± 12.5 (p = .14) in the active arm and from 17.6 ± 6.6 to 19.1 ± 8.1 (p = .56) in the control arm, respectively.Word Fluency Test. Both treated groups performed similarly with an average score change from 20.3 ± 7.3 to 22.9 ± 7.6 (p < .001), Trial 5 by 31.2% (p = .001), Trial 1-5 Sum by 33.9% (p < .001), and Trial 7 by 47.7% (p = .002). On other hand, sham treated patients did not demonstrate statistically significant results in the end of the treatment. In most subtests, response to sham treatment had no change in performance  or insignificant trend  of improvement . Treatment with active device resulted in statistically significant positive cognitive improvement in patients’ performance during the course of treatment. Immediate A.V.L.T.-1 test resulted in improvement of evaluated performance for Trial 1 by 48.6% (rovement .p = .015) over the course of treatment. Significant observation was also noted for recognition subtest with performance improvement by 14.5% (p = .05) compared to score at the beginning of the treatment. However, sham treated patients did not demonstrate significant improvements on aforementioned subtests . Delayed (30 min) recall and recognition subtests were evaluated in patients from both groups. Significant improvement was noted in the active group in delayed recall subtest with an average score improvement by 2.2 (63.5%) points increasing from 3.4 ± 2.9 to 5.6 ± 4.4  with ease in the friendly environment of home and minimal instructions. This approach indicates that such technology can be used for a long period of time remotely by self-administration or with the help of a family or friend caregiver. A daily log was provided to patients and caregivers to impose treatment adherence and compliance. Portable and cushioned cases were provided along with the helmet device to ensure safe storage of the device and offer portability, increasing treatment accessibility and minimizing the potential conflict between treatment schedule and unexpected life events . These positive aspects of the treatment were represented in feedback from participants and their family caregivers. In comparison, other commonly used medical devices could be either invasive, such as a cardiac pacemaker, or require long hours continuous use causing discomfort, such as a continuous positive airway pressure (CPAP) machine. This demonstrates the feasibility and acceptability of a light emitting device, suggesting that the device can be a part of normal living in patients with dementia who need prolonged, continuous, and uninterrupted treatment without compromising quality of life.Our study demonstrated safety of 1060-1080nm emitting spectrum, low power near infrared light used twice daily in this trial. This study is a second tNIR light treatment trial completed in Baylor Scott and White Health following a smaller recent pilot safety study using same technology and safety protocol, which was successfully conducted and completed in collaboration with Dr. Marvin Berman and his group from Quiet Mind Foundation, Elkins Park, PA, USA . The twoAs we look at the subtests from different individuals, the outcomes are even more exciting over only 8-week of active treatment: a participant with moderate dementia (MMSE = 16 prior to treatment) achieved a 75% improvement (MMSE = 28 at the end of the treatment); another patient had 80% improvement in the clock drawing tests (Schulman method scoring); two patients had 60% and 80% performance improvement in the clock copying test; two patients had 12 points  and 13 points  improvement in logical memory immediate and delayed subtests; executive function has improved by up to 73% in some patients evidenced by both trail making test A and B, and verbal learning and memory tests have improved by up to 35% as shown in the active verbal learning immediate and delayed tests. In general, across multiple subtests, patients in the group treated with active tNIR light emitting device demonstrated considerable improvement while patients in the sham treated group did not. These observed positive outcomes suggest beneficial effects of tNIR light stimulation on degenerative progression of neuropathological conditions associated with dementia. In sham treated group, natural progression of disease during the course of the study contributed to no positive change or decline in performance outcomes.In the LMT-I, LMT-II, A.V.L.T.-1, A.V.L.T.-2 subtests and trail making test A, the pre-treatment mean values in the sham treated group appeared to be lower than pre-treatment mean values in the active group. However, the pre-treatment means values for sham treated group were higher in MMSE, CDT, CCT, DSB and trail making test B assessments. This could be attributed to the difference in mean age of study population in active (72.4 ± 8.2 y.o.) and sham (77.8 ± 5.2 y.o.) groups, to privilege of male population in active (59%) versus sham (53%), to relatively small sample size in the control group (n=17) versus active group (n=40), as well as explained by the underlying neuropathology. In addition, inherent ceiling and flooring effects could limit the reliability of these subtests. Nonetheless, further investigations with larger sample size, contribution of sex, age and causing neuropathology are warranted.Our study also demonstrated improvement of sleep-in active group compared to feedback from patients and family caregivers in the sham group. Although without any specific measurement, it carries scientific merit and should be integrated into future studies given the important role of long and uninterrupted sleep-in dementia and elderly population, as sleep disturbance has been well demonstrated in dementia patients . CaregivRecent pre-clinical and clinical studies using tNIR light stimulation reported promising improvements in treatment of traumatic brain injury (TBI), Parkinson’s disease (PD), and Alzheimer’s disease (AD). Below we discuss latest findings and mechanisms of PBM in treatment of neuropathological conditions.A few studies have investigated the application of PBM in the treatment of TBIs. Some demonstrated successful treatment of TBI associated symptoms using red light stimulation , 30. As in-vitro, in-vivo and growing number of recent human clinical trials in this field [With respect to TBI triggered inflammatory chain reaction and initiated generalized immune response, we support the idea that aforementioned pathological processes can be effectively addressed by implementing tNIR light therapy. This approach is supported by literature to be quite effective in acute and chronic TBI treatment, considering general safety and demonstrated efficacy of tNIR light stimulation is field , 30. Oneis field .Parkinson’s disease is a chronic CNS pathology caused by slow degeneration of substantia nigra (SN) pars compacta dopaminergic neurons. PD has various clinical presentations with motor and non-motor characteristics depending on the stage of disease. The most common symptoms are resting tremor, slowness of movements, rigidity of muscles, balance impairment, dementia, anxiety, depression, hallucinations and delusions. Recently, several studies have investigated the effects of tNIR light and PBM in treatment of PD using mice and monkey models -38. MostIn this study, we have observed reduction of tremor with noticeable improvement of balance in patients with PD associated dementia. This could be consolidated with the improvement of micro-tremor observed during clock drawing test in this study . More clAlzheimer’s disease is the most common neurodegenerative pathology of CNS with progressive memory loss and cognitive function impairment . In AD, Patient with mild dementia may have less room for improvement compared to patients diagnosed with moderate dementia. Additionally, some tests may not be powerful enough to detect fewer substantial changes. Therefore, robust changes in individual performances can be obscured by those with minimal changes and vice versa, when analyzing combined datasets from different stages of disease . This can potentially be addressed by stratifying patients’ data based on severity of dementia. Results of stratified patients’ data by stage of disease may represent a full potential of therapeutic range for each patient group.Transcranial applications in this study demonstrated safe and convenient approach to deliver NIR light to the brain without local or systemic side effects and adverse reactions. Treatment protocol designed for this trial was simple and successfully used at the convenience of home by elderly population. Observed positive cognitive, executive, and mood changes can benefit patients with dementia by improving quality of life and self-independence in daily lives, and thus helping their immediate family caregivers by decreasing their burden. More studies are necessary to look into family caregivers’ burden and to ensure reproducibility of positive findings.


Chronic Kidney Disease is a worldwide public health problem that affects millions of people from all racial and ethnic groups. Identification of a Chronic Kidney Disease is a major risk factor for cardiovascular morbidity and mortality and is attributed to hyperuricemia. Evidences show that high serum uric acid contribute directly to glomerulosclerosis, interstitial fibrosis and atherosclerosis that correction of hyperuricemia associated with Chronic Kidney Disease will slow the progression of chronic renal failure. This study is done to correlate between serum uric acid level and estimated glomerular filtration rate in chronic kidney disease patients.A hospital based cross-sectional study on chronic kidney disease patients including 57 patients on conservative treatment attending Bir Hospital with diagnosis of chronic kidney disease was performed. Detailed clinical history, examination and investigations including uric acid were done. Chronic Kidney Disease staging was done according to estimated glomerular filtration rate estimated by Cockcroft-Gault equation. Prevalence rate of hyperuricemia in Chronic Kidney Disease and its stages were calculated and compared with each other.A total of 57 Chronic Kidney Disease cases were enrolled, with male to female ratio of 2:1 and mean age 51.63±17.75 years. Hyperuricemia was present in 55 (96.49%) of study population. Though prevalence of hyperuricemia increased with Chronic Kidney Disease stage, there was no significant difference in mean value of uric acid in different stages. Hyperuricemia and stages of Chronic Kidney Disease had negative correlation which was statistically significant.Hyperuricemia is highly prevalent among Chronic Kidney Disease patients with conservative management. The severity of hyperuricemia increases as Chronic Kidney Disease stage progresses. Kidney damage refers to a broad range of abnormalities of kidney resulting in decline of excretory, endocrine and metabolic functions. KDIGO refers to a eGFR<60 ml/min/1.73m4 Evidence reported on an association of uric acid with hypertension and renal disease.5 It is a potent antioxidant outside cell, pro-oxidant inside cell and induce stimulation of NADPH oxidases with mitochondrial dysfunction.6 It leads to arteriosclerosis, glomerular hypertension and glomerulosclerosis and progress CKD.7Chronic hyperuricemia is strongly associated with chronic tubulointerstital disease and decreased renal function.This study is done to know the prevalence of high serum uric acid level in chronic renal disease and to correlate between severity of the renal disease and serum uric acid level.It is a hospital based cross-sectional study. It comprises 57 patients with CKD on conservative management who attended Department of Nephrology, Bir Hospital from May 2015 to April 2016. The study was performed after approval from the Institutional Review Board at our hospital and taking written, informed consent from all the patients. All cases of CKD stage- 3,4,5 who were on conservative management were included. Excluded cases were: 1. Renal transplant patients, 2. Patients on uric acid lowering drugs, 3. CKD patients on hemodialysis and 4. Patients not willing to give consent.Non-random sampling technique was applied. Patients admitted and attending outpatient department in hospital with clinical, biochemical and sonographic evidence of CKD were taken. Sample size was calculated using following formula:n= required sample sizeZa= z deviated corresponding to desired reliability level (1.96)8P= estimated proportion in the population (10.6%)Q= 100-P (if P is in %)d = maximum tolerable error (10%)WhereCKD staging is done as following  . A detaData was tabulated and interim analysis was performed after the reports of 10 cases are available. All the analysis were carried out using standard statistical software SPSS 20 version and Microsoft Excel. Results are presented in tables, graphs and diagrams. Results were expressed as mean±SD. Prevalence rate of hyperuricemia in CKD and its stages were calculated and compared with each other. Statistical analyses included the unpaired t-test (for continuous measures) and the Chi-square tests for categorical assessed. For the purpose of this study a 95% confidence interval was accepted and a P<0.05 was taken as significant.Out of 57 patients, 38 were male and 19 were female. The mean age for the total number of patients was 51.63 years. Hyperuricemia was present in 55 (96.49%) of the patients .The mean uric acid level in men was 9.38±1.403 mg/ dl and mean uric acid level in female population was 8.86±1.13 mg/dl .The mean uric acid level in stage 3 CKD in female population was 12 mg/dl and in men population was 8.67 mg/dl. In stage 4 CKD mean uric acid level in female was 8.76 mg/dl and in men was 9.24 mg/dl and in stage 5 CKD mean uric acid level in female was 8.55 mg/dl and in men was 10.15 mg/dl .Of the study population 15 (26.31%) had hypertension, 6 (10.52%) had hypertension and DM, 18 (31.6%) had hypertension and smoking, 2 (3.5%) had DM, 9 (15.8%) had hypertension, DM and smoking and 2 (3.5%) were only smokers .2. The mean serum uric acid of the study participants was 6.5±1.9 mg/dl.9This study has evaluated prevalence of hyperuricemia in Chronic Kidney Disease. Patients in a cohort of 57 subjects, out of which 38 patients (66.7%) were male and 19 patients (33.3%) were females. In this study, hyperuricemia was present in 96.49% of study population while 3.51% has normal serum uric acid. The mean serum uric acid in the study population was 9.213 mg/dl. Similar study done by Jolly ES et al. in which a 1078 participants was analysed in Genetics of Coronary Artery Disease in Alaska Natives (GOCADAN) study in which the participants were followed for 4 years. Sixty three (13%) of the participants had CKD. The mean age of the participants having CKD was 63 years. Forty seven (63%) of them had HTN and 7 (9%) had DM. BMI of the CKD patients was 28.5 kg/m10In the study done by Paudel YP et al. at Kist Medical College and Teaching Hospital in Kathmandu involving 150 CKD patients, found 57% male, 43% female, mean age was 48.62±18.09 years. The mean uric acid level was 5.96±2.22mg/dl. In this study, the prevalence of hyperuricemia in various stage was CKD stage 3–24.6%, stage 4–40.4% and stage 5–35% study population. There is no significant difference in the mean value of serum uric acid in the various stages of CKD.11Similar study done by Adejumo OA et al, prevalence of hyperuricemia in stage 3 CKD patients was 33.4%, stage 4–40.8%and stage 5–21.7%. In this study, mean creatinine level of the patient was 4.1298 mg/ dl. The mean systolic BP was 156.3860 mm of Hg and mean diastolic BP was 93.7544 mm of Hg. Out of 120 patients, 57 of the patients had high serum uric acid level. The prevalence of hyperuricemia was 47.5%.12 The association was more pronounced in hypertensive subjects compared to normotensive individuals. In this study the most common cause for CKD was hypertension 57.89% of studied population had HTN. Hypertension and DM both were present on 27.31% of the patient. DM alone was present on 3.5% of the patients. The remaining 12.28% of the patient had other causes for CKD. In the study done by Adejumo OA et al. involving 120 patients the causes of CKD were that 37.8% of the patient had hypertensive nephropathy, 30% had diabetic nephropathy, 27.5% had chronic glomerulonephritis and 5.8% had other causes for CKD.Most of the patients in our study were hypertensive, so it can be concluded that hyperuricemia may lead to the progession of CKD faster in hypertensive patients. Study done by Sedaghat S et al. in a prospective cohort showed that high levels of serum uric acid are associated with faster decline in eGFR and increased incidence of CKD.This study showed that hyperuricemia is highly prevalent among CKD patients with conservative management. Though this study showed increasing prevalence of hyperuricemia with increasing stage of CKD (stage III to V), the mean value of uric acid did not show significant difference among different stages. Further studies without above mentioned limitation and including larger group are needed.None.


The first aliquot was not subjected to any intervention. The second, third and fourth aliquots were passed through a fine needle 2, 4 and 6 times, respectively. Complete blood count was performed by multi-angle polarized scatter separation technology and haemolysis index (HI) was assessed from the plasma samples separated by centrifugation. Five groups were formed according to the HI values. The percentage biases between the results of non-haemolysed and haemolysed groups were compared with the desirable bias limits from The European Federation of Clinical Chemistry and Laboratory Medicine database and reference change values (RCVs).In groups 1 to 4, the effects of haemolysis on CBC parameters were acceptable comparing to the analytical bias except for lymphocytes (7.26%-7.42%), MCH (2.59%), and MCHC (0.47%-2.81%). Results of group 5 (gross haemolysis) showed decreases in HCT(- 4.56%), RBC (- 4.07%) count and increase in lymphocyte (11.60%) count higher than the analytical performance specifications. Moreover, variations in MCH (4.65%) and MCHC (5.24%) were exceeding the RCVs.Gross haemolysis (haemoglobin concentration > 10 g/L) is likely to produce unreliable CBC results on non-pathological samples. Further studies including pathological specimens are needed. In vitro haemolysis is one of the most common preanalytical error affecting many test results. Release of haemoglobin (Hb) and the other intracellular contents secondary to impaired membrane integrity of red blood cells (RBC) may cause artifactually increase or decrease results. Inappropriate techniques for collection, transport and processing of samples are the major causes of in vitro haemolysis. Haemolysis may simply be detected by visual inspection of the serum or plasma component of the sample when it contains enough quantity of Hb to notice. More recently, automated analysers can quantify cell-free Hb by photometric measurement at different wavelengths following centrifugation  as whole blood specimens are used. For this reason concomitant serum or plasma samples may be evaluated for haemolysis or dramatically increased mean corpuscular haemoglobin concentration (MCHC) may be used as warning for the prevention of unnoticed haemolysis  employs optical scatter and fluorescence technologies as well as cyanide-free absorption haemoglobinometry to perform CBC .Blood samples were drawn into two 3.0 mL evacuated tubes containing K3- ethylenediaminetetraacetic acid (EDTA) (Ref number:IST413)  and mixed gently immediately after collection. Samples were portioned in order to obtain four aliquots of 1.5 mL from each subject subsequently. The first aliquot did not undergo any intervention while the second, third and fourth aliquots were passed through a small calibre needle  2, 4 and 6 times, respectively in order to produce scalar amounts of haemolysis according to the Dimeski method . The HI had 7 semi-quantitative values (0 – 6) corresponding to Hb concentrations of ≤ 0.10, 0.11-1.30, 1.31-2.49, 2.50-4.99, 5.00-7.49, 7.50-9.99 and≥ 10 g/L, respectively. All samples were grouped according to HI values.W) and analytical variation (CVA) with the formula: RCV = ± 1.96 x (CVA2 + CVW2) ½ .The Bland-Altman plot analysis is used to asses the biases between the results of paired groups. Effect of haemolysis interference was evaluated according to the analytical performance specifications (APS) for desirable bias as obtained from the European Federation of Clinical Chemistry and Laboratory Medicine (EFLM) European Biological Variation Study . The samples were presented as 5 groups according to the HI values; Group 1: HI 1 , Group 2: HI 2 , Group 3: HI 3 , Group 4: HI 4 , and group 5: HI 6 . There were no samples with HI 5 (Hb concentration 7.50–10.0 g/L). As the degree of haemolysis was different among the samples due to mechanical trauma and to determine the relative percent bias accurately HI 0 groups were paired separately for each group from the non-haemolysed portions (N = 25). Bias percentages and confidence intervals determined for each paired group are presented in In group 1 there was no significant difference in any of the measured parameters.Results of group 2 showed that at HI 2 all the parameters, with the exception of MCHC, had alterations meeting the APSs. Percent bias of MCHC [0.47 (- 0.4 to 1.33)] was slightly higher than the error limit (± 0.4%) defined according to EFLM database but was still acceptable when evaluated according to RCV (4.12%).In group 3, lymphocyte count and MCHC results had percent bias values: 7.26 (2.84 to 11.68) and 0.73 (- 0.13 to 1.58) above the APS related error limits , but they were acceptable in terms of RCVs .Group 4 demonstrated that for the lymphocyte count, MCH and MCHC parameters, the percent bias 7.42 (1.45 to 9.08), 2.59 (1.37 to 3.80) and 2.81 (1.61 to 4.03) were exceeding the analytical limits  but all values were acceptable according to RCVs.For the grossly haemolysed group 5, lymphocyte counts had percent bias of 11.60 (5.57 to 17.62) exceeding the analytically desirable limit (± 6.3%) but were still lower than RCV (31%). RBC count and HCT were significantly decreased in grossly haemolysed (HI 6) samples as compared with non-haemolysed ones. The percent bias results of RBC count and HCT (- 4.07 (- 5.52 to - 2.62) and - 4.56 (- 6.13 to - 3.00), respectively) were higher than the analytical bias goals  but under the RCVs (7.46% and 8.11%). However, the results of MCH and MCHC in this group were affected significantly according to both of the error limits.S. On the other hand, gross haemolysis (> 10.00 g/L Hb concentration) led to decreases in HCT, RBC count and an increase in lymphocyte count higher than the desirable bias. Moreover, variations in MCH and MCHC were exceeding the RCVs. Investigation of the impact of haemolysis on CBC is an important requirement. To our knowledge, this is the first study assessing the haemolysis interference in relation to both analytically and clinically significant error limits on the Abbott Alinity hq system.We observed that the effect of 0.11 to 7.49 g/L Hb concentration on CBC parameters were acceptable comparing to the analytical biases with the exceptions of lymphocytes, MCH, and MCHC. In addition, all the parameters were within RCVet al. indicated that haemolysis caused 7.3% (- 34.2% to 48.7%) and 42.6% (- 16.8% to 101.9%) bias for PLT count at mild and high degree, respectively  with the exception of MCHC. This was supporting the findings of De Jonge In the Hb analysis, RBCs were lysed using a reagent and released Hb was measured with absorption photometry. Haemoglobin measurement was not affected from haemolysis because the method was not specific for intracellular or free Hb. This finding was concordant with the previous studies (The results of this study showed that MCV and RDW (%) were not interfered significantly with haemolysis comparing to the desirable analytical and clinical bias criteria. This finding was incompatible with the previous studies . In addition, the different reactions of the erythrocyte membranes of different samples to mechanical trauma might have been generalized with a wider study group. The second limitation of our study is that we could not find the possibility to evaluate haemolysis in pathological specimens.In conclusion, gross haemolysis (Hb concentration > 10 g/L) is likely to produce unreliable CBC results in non-pathological samples. Further studies including pathological specimens are needed.


Human papillomaviruses (HPV) are non-enveloped DNA viruses infecting cutaneous and mucosal squamous epithelia. Sexually transmitted HPV-types that are carcinogenic to humans such as HPV16 can induce cervical and other anogenital cancers. Virus transmission through fomites such as inadequately disinfected gynecological equipment is a further potential transmission route. Since HPV cannot be easily grown in cell culture, polyomavirus SV40 has been used as a surrogate virus when testing the virucidal activity of chemical disinfectants. So far, studies that have compared the virucidal activity of different disinfectants against HPV and SV40 are lacking. Here, we evaluated the susceptibility of HPV16 pseudovirus and SV40 to seven active biocidal substances using quantitative suspension tests. Ethanol, glutaraldehyde (GTA), dodecyldipropylentriamin (DPTA), and ortho-phthalaldehydes (OPA) were able to reduce the infectivity of HPV16 pseudovirus >99.99% after 5 min. In contrast, isopropanol, peracetic acid (PAA), and quaternary ammonium compounds with alkylamines (QAC) only led to a slight or no reduction in infectivity. Concerning SV40, only GTA (60 min contact time), PAA, and OPA had virus-inactivating effects. In conclusion, the virucidal activity of three out of seven disinfectants tested was different for HPV16 pseudovirus and SV40. In this study, SV40 was shown to be a reliable surrogate virus for HPV when testing isopropanol-, GTA-, QAC-, and OPA-based disinfectants. Anogenital human papillomaviruses (HPV) are transmitted by sexual contact and HPV in risk group 1, which are carcinogenic to humans, can cause high-grade cervical, vaginal, vulvar, and anal intraepithelial neoplasia and the respective anogenital cancers ,3. HowevThe HPV life cycle is tightly linked to the host cell differentiation, which makes it difficult to grow HPV in standard cell cultures. To overcome this difficulty, different cell culture systems to produce recombinant viral particles have been developed. So called quasiviruses have a capsid consisting of codon optimized L1 and L2 proteins and contain an HPV genome. Pseudoviruses also have codon optimized L1 and L2 capsid proteins but, in contrast to quasiviruses, encapsidate a GFP reporter plasmid (pseudogenome) that can easily be tracked microscopically in cellular infectivity assays ,13,14,15In the past, both HPV and the polyomavirus simian virus 40 (SV40) were assigned to the family Papovaviridae, a now abolished family, which was split into the two families Papillomaviridae and Polyomaviridae in 1999 . Based ohttps://vah-online.de/en/dmk-working-groups, (accessed on 5 June 2021)). The substances tested are listed in In order to prevent intra-laboratory variations, calibrated solutions of disinfectants used in the HPV16 pseudovirus and SV40 assays were provided by the task force for interlaboratory surveys  of the Association of Applied Hygiene (VAH) c/o Institute for Hygiene and Public Health, University Hospital Bonn, Bonn, Germany  and quaternary ammonium compounds (QAC) with alkylamines are commonly used as disinfectants in clinical settings, and peracetic acid (PAA), dodecyldipropylentriamin (DPTA), and orthophtalic acid (OPA) are used for the disinfection of medical instruments and surfaces.2 atmosphere. 293TT cells replicate plasmids with the SV40 origin of replication to a very high copy number, because they express the SV40 T-antigens in their nucleus, enabling replication of the HPV16 pseudovirus genome (The human embryonic kidney cell line 293TT was maintained in Dulbecco’s modified Eagle’s medium (DMEM)  with 10% fetal calf serum (FCS) (Life Technologies), 1× penicillin-streptomycin  (Life Technologies) and supplemented with 250 µg/mL hygromycin B . Cell cultures were incubated at 37 °C in a humidified 6% COs genome  11]..2 atmospg for 5 min. The final wash step was conducted with PBS containing 0.8 M NaCl. Pseudovirus containing supernatants were loaded onto an Optiprep  gradient  and centrifuged for 3.5 h at 16 °C at 50,000 rpm  in an SW55ti rotor. Virus titers were determined by incubating 293TT cells (1 × 104 cells/well in 96-well plates) with a 1:10 dilution series of HPV16-pseudovirus fractions (n = 8 repeats for each dilution). After 48 h, cells were analyzed for GFP expression by fluorescence microscopy /Robert Koch Institute (RKI) . The int2 for 48 h).The remaining virus titer was again calculated using the Spearman and Kärber method ,20. The 2 atmosphere. The cells were fed with Dulbecco’s minimum essential medium (D-MEM) supplemented with heat-inactivated 10% FCS and non-essential amino acids. For virus cultivation, confluent monolayers with a maximum age of two days were used and infected with SV40. Cells showing a cytopathic effect were harvested and virus was isolated by one freeze and thaw cycle. Cell debris was separated by low-speed centrifugation at 400× g for 10 min. Aliquots of the virus suspension were stored at −70 °C.The SV40 stock virus suspension was produced according to the DVV/RKI Guidelines. Polyomavirus Simian virus 40 (SV40) strain 777 and CV-1 cells, an immortalized rhesus monkey kidney cell line for virus cultivation and suspension tests, were kindly provided by the DVV virus bank . CV-1 cells were cultivated at 37 °C in a humidified 5% CO7−108 tissue culture infectious dose, 50% (TCID50/mL)) and one part by volume of the interfering substance  was mixed with eight parts by volume of the disinfectant at 20 °C ± 2 °C. Infectivity was determined in microtiter plates by means of end point dilution titration. At the end of the chosen contact time (3 cells per well), 100 μL from each dilution was placed. Cultures were observed for cytopathic effects (CPE) for at least 14 and up to 21 days after exposure. All tests were conducted in three independent test runs that were performed on different days in two independent laboratories, respectively. Virus controls were evaluated after the longest contact time. The RF was calculated as the difference between the remaining SV40 virus titer after exposure to disinfectant and unexposed SV40. Cytotoxicity was determined by serial tenfold dilution with sterile water of standardized hardness instead of test virus suspension.Tests were performed in two of our laboratories (Stuttgart and Bremen) according to the Guidelines of the German Association for the Control of Virus Diseases (DVV)/Robert Koch Institute (RKI) . The intact time , the rea50/mL with standard deviation. Titer reduction is presented as the difference between the virus titer after contact time with the disinfectant and the titer of unexposed virus. This difference is given as titer reduction factor ((RF), log10 reduction of viral titer) with standard deviation. A reduction in viral titers of ≥4 log10 steps (inactivation ≥ 99.99%) was regarded as proof of virucidal activity.The virus titers were determined using the Spearman and Kärber method ,20 and eIn this study, we evaluated the susceptibility of HPV16 pseudovirus and SV40 to different disinfectants in order to determine whether SV40 is an appropriate surrogate virus for HPV in the context of studies on chemical disinfectants. To this end, we compared HPV16 pseudovirus and SV40 with regard to their susceptibility to ethanol, isopropanol, and five further disinfectants .10 steps, it is defined as virucidal activity. Disinfectants based on GTA, DPTA, and OPA were also able to reduce virus titers by >99.99% in all concentrations and contact times tested (Experiments with the HPV16 pseudovirus showed that longer contact times (12 min and 60 min) did not have a statistically significant impact on infectivity compared to 5 min A. From ts tested . On the s tested A.10 steps under all conditions tested and thus were not considered to exhibit virucidal activity. The same was true for GTA 0.05% and PAA 0.005% at a contact time of 5 min, respectively. However, SV40 incubated for 60 min with disinfectants based on 0.03% or 0.05% GTA were shown to reduce infectivity ≥4 log10 steps. The same was true for 0.05% PAA (contact time 5 min) and 0.55% OPA (contact time 5 min and 12 min)  B.Of the disinfectants tested, only GTA 0.03% and 0.05% at a contact time of 60 min and OPA 0.55% at contact times of 5 and 12 min, respectively, showed virucidal activity against both HPV16 pseudovirus and SV40. Ethanol , DPTA 0.5% (5 min and 60 min), and GTA 0.05% at a contact time of 5 min were only effective against HPV16 pseudovirus, while PAA 0.05% at a contact time of 5 min only inactivated SV40. Isopropanol , PAA 0.005% (5 min), and QAC 0.5% (5 min and 60 min) were ineffective against both HPV16 pseudovirus and SV40 A,B.Up until now, European Guidelines lack a suitable test virus to validate the efficacy of disinfectants against HPV . In contIn a previous study by Meyers et al. 2014), HPV16 native viruses, isolated from organotypic culture systems and quasivirus were analyzed for their resistance against chemical disinfectants. The authors tested ethanol and isopropanol and even after a contact time of 45 min, no reduction in infectivity of both virus stocks was observed with both 70% and 95% ethanol . In thei, HPV16 n10 reduction in SV40 virus titer was measured with disinfectants based on DPTA (contact time 60 min). This indicates that SV40 could be susceptible to this substance. However, the predefined requirement for virucidal activity was not met by DPTA for SV40. Interestingly, significant virucidal activity against SV40 was observed for disinfectants based on OPA (contact time 5 min) and GTA (contact time 60 min). In addition to ethanol, we also detected a strong reduction in HPV16 pseudovirus titer after incubation with disinfectants based on GTA, DPTA, or OPA. The virucidal activity of OPA found in our study is in accordance with Egawa et al. (2021), who have recently shown that OPA inactivates HPV16 pseudovirus and HPV18 native virions . In the When comparing the results for HPV16 pseudovirus and SV40, we found that both were resistant against 60% or 70% isopropanol and 0.5% QAC+ alkylamines. In contrast, 60% and 70% ethanol could inactivate HPV16 pseudovirus after a contact time of 5 min, but not SV40. With respect to ethanol-based disinfectants, SV40, being more resistant to ethanol, can be seen to cover HPV16. The use of SV40 as a surrogate for HPV16 in the case of PAA needs to be seen with caution as we could show that 0.05% PAA is active only against SV40, but not against HPV16 pseudovirus. Here, either a more resistant virus  to clarify the partly contradictory results of published studies.