ASA 1 individuals  had decreased odds of EIF, and implants in those using anticoagulants  had increased odds of EIF. At the patient level, the odds of EIF in ASA 3  and IHD  individuals decreased. In AF/VF  individuals, EIF odds increased. Conclusions: Within the limitations of the present study, the use of AC is significantly associated with an increased likelihood of EIF: the OR was 2.64. Future research is required to validate and examine the prospective impact of AC on the osseointegration phenomena.Background: Anticoagulants (AC) are among the most often prescribed drugs in the world. Data regarding ACs’ effect on the osseointegration of dental implants is lacking. Purpose: The aim of the present retrospective cohort study was to evaluate the effect of anticoagulants (AC) on early implant failure (EIF). The null hypothesis was that the use of AC increases the incidence of EIF. Materials and Methods: The research included 687 patients who underwent 2971 dental implant placements in the department of oral and maxillofacial surgery in Rabin medical center, Beilinson hospital, by specialists in oral and maxillofacial surgery. The study group comprised 173 (25.2%) patients and 708 (23.8%) implants using AC. The rest of the cohort served as a control. A structured form was used to collect data at patient and implant level. EIF was defined as implant failure within a period of up to 12 months from loading. EIF was the primary outcome parameter. A logistic regression model was used to predict EIF. Results: Implants placed in individuals ≥ 80 (odds ratio (OR) = 0.34,  Implant dentistry is a safe and current therapeutic option for patients who are totally or partially edentulous. Research into adverse events endangering osseointegration has appeared more and more in the existing literature . Dental The time of implant failure occurrence is used to categorize failures . Early iReports on the impact of systemic disorders on implant therapy may be found in the dental literature. Still, many oral and systemic diseases have unclear effects on osseointegration .One of the most prevalent complications associated with endosseous implants is hemorrhage . AnticoaExtensive research has been conducted on the clinical effects of VKAs on human bone metabolism. There is evidence that VKAs affect bone metabolism. Long-term usage of oral AC has been related with an increased risk of osteoporosis. As a VKA, warfarin has a detrimental effect on bone health. Vitamin K is essential for osteocalcin’s posttranslational glutamination, the primary non-collagenous bone matrix protein. Dietary limitations typically undertaken by patients receiving VKAs are one additional potential indirect mechanism of bone deterioration ,7.There has only been a limited amount of clinical research on the effects of DOACs on bone since they are still relatively new to clinical practice. The possible effect that DOACs may have on bone metabolism have been poorly investigated so far ,9. In 20The aim of the present retrospective cohort study was to evaluate the effect of anticoagulants (AC) on EIF. The null hypothesis was that use of AC increases the incidence of EIF.The research included 687 patients who underwent 2971 dental implant placements in the department of oral and maxillofacial surgery in Rabin medical center, Beilinson hospital, by specialists in oral and maxillofacial surgery. Early implant failure (EIF) was defined as the removal of the implant from the oral cavity within a period of up to 12 months from the date of its loading in the oral cavity.Patients who underwent dental implant placement in the department of oral and maxillofacial surgery in Beilinson hospital.Follow-up, ≥12 months from the date of implant loading.Each patient had a CT scan before the transplant.Follow-up included X-rays (panoramic), and clinical examination.Inclusion criteria:Follow-up of at least <1 year from loading.Severe systemic diseases that include immunosuppression.Patients who underwent head and neck radiation ≥ 5000 rad.Exclusion criteria:Age Group—≤65 or 66–79.9 or ≥80.Physical status—ASA 1/ASA 2/ASA 3 .Smoking status—No/Yes.Use of AC.Systemic diseases—HTN; DM; CVA; hyperlipidemia.Implant brand—Ditron Dental/MIS/Zimmer Dental.Bone augmentation—Pristine/Augmented.Implant location—maxilla/mandible; anterior/premolar/molar.Implant dimensions—length, diameter.EIF—primary outcome parameter.Variables collected at implant level included:Gender—male/female.Age Group—≤65 or 66–79.9 or ≥80.Physical status—ASA 1/ASA 2/ASA 3.Smoking status—No/Yes.Use of AC.Systemic diseases—IHD; AF/VF.Bone augmentation—Pristine/Augmented.Total implant number placed per individual.EIF—primary outcome parameter.Variables collected at patient level included:p-value < 0.05 was considered statistically significant.The software SPSS version 25 was used for data analysis. Tests conducted were as follows: Chi-square (The cohort (n = 687) comprised 61.7% males and 38.3% females. Ages (years) were: 58.2% ≤65 years, 32.5% 66–79.9 years, and the remaining 9.3%, ≥80 years or older. Physical status was: 32.5%—ASA 1; 35.4%—ASA 2, and the remaining 32.2%—ASA 3. A quarter (173/25.2%) used anticoagulants. Systemic diseases included: 12.2%—ischemic heart disease (IHD); and 8.9%—atrial/ventricular fibrillation (AF/VF). Smokers made up 5.4%. Implant brands were: 12.5%—Ditron Dental; 27.4%—MIS; and 60.1%—Zimmer Dental. More than half (55.7%) of the individuals required bone augmentation for implant placement. The EIF cohort at patient level was 15.1%.All data were likewise analyzed at implant level (n = 2971). Ages were 57.7% ≤65 years, 34% 66–79.9, and the remining 8.3% ≥80 or older. Physical status was 27.3%—ASA 1, 39.6%—ASA 2, and 33%—ASA 3. A quarter (708) of the implants (23.8%) were inserted in individuals using anticoagulants. Implants inserted in individuals with systemic diseases included: hypertension (HTN)—29%; hyperlipidemia—23.6%; DM—14.9%; and cerebrovascular accident (CVA)—4.6%. Implants inserted in smokers made up 6.6%. Implant brands were: 13.6%—Ditron, 26.3%—Mis, and 60.1%—Zimmer. Most of the implants (61.2%) required augmentation. The implant location was: 15.3% anterior maxilla, 18.5% premolar maxilla, 15% posterior maxilla, 13.7% anterior mandible, 17% premolar mandible, and 20.5% were for posterior mandible. Lastly, EIF was recorded for 3.8% of the implants. For a complete description, see At implant level , signifiSmokers were more likely to use AC (8.6% vs. 5.9%) (A significant difference was found for the ASA group (p = 0.05). EIF was more likely to occur in those using AC (5.5% vs. 3.3%) (Those with implants in the anterior mandible were more likely to use AC (18.9% vs. 12%)  = 33.07, p = 0.005), while it explains about 9.6% of total variance in failure. The model is well fitted to the data (χ2(8) = 2.78, p = 0.95), while it classifies about 96.3% of the total observations.A logistic regression model at the implant level showed that the independent variables significantly predict failure (p = 0.05), and ASA 2/3 vs. ASA 1 individuals  had decreased odds of EIF and implants in those using anticoagulants  had increased odds of EIF. For complete regression coefficients, see Implants placed in individuals ≥80  = 27.12, p = 0.001), while they explain about 7% of total variance in failure. The model does not fit to the data well (χ2(6) = 8.31, p = 0.22), while it classifies about 85.0% of the total observations.A logistic regression model at the patient level showed that the independent variables significantly predict failure (p = 0.02) and IHD  patients are decreased. In patients with AF/VF , the EIF odds are increased. No statistically significant difference in EIF odds was found for those using AC (p = 0.87) while controlling for the other variables. For complete regression coefficients, see At patient level the odds of EIF in ASA 3  score (ASA 2 or 3) had lower odds of implant failure than those with an ASA 1 score. These data imply that, depending on the ASA classification, there might be protective factors or other mechanisms that influence the probability of EIF. The treatment and management of patients with a higher ASA score may have been more thorough, with a greater emphasis on avoiding risks and optimizing outcomes, which may explain this finding . FurtherIt was found that EIF odds for patients with IHD were lower. Patients with AF/VF had increased EIF. Patients with serious medical conditions are often managed with medications and lifestyle changes to reduce the risk of complications. These interventions may also help to reduce the risk of EIF. It is probable that this is reflected in the data for patients with IHD. It is also worth noting that there may be other factors that contribute to the relationship between IHD and EIF, or between AF/VF EIF. To better understand these relationships and to identify the most effective strategies for reducing implant failure in patients with different medical conditions, it will be important to conduct additional research and to consider a wider range of factors that may be influencing the outcome.One of this study’s limitations is not measuring bone densitometry due to its retrospective nature. Future studies should take this factor into account. There is also the possibility of bias: for patients who take oral anticoagulants, this is usually due to having a cardiac arrhythmia such as atrial fibrillation. This means older age, higher rates of hypertension, and possibly, smoking, factors that also affect bone remodeling.Osseointegration was compared between 80 implants with hydrophilic SLActive and hydrophobic SLA surface in 20 anticoagulated patients. A “split-mouth” study design was used. After one year, 100% implant survival and success rates were observed .A retrospective, study  assessed anticoagulant drugs’ effect. A 4.4% lower survival rate was reported, and the odds ratio of failure was 28.2. It was concluded that anticoagulants increase the risk of implant failure .A recent review revealed that non-steroidal anti-inflammatory drugs, glucocorticoids, proton pump inhibitors, selective serotonin reuptake inhibitors, anticoagulants, metformin, and chemotherapeutic agents may jeopardize osseointegration and lead to implant failure. However, validation is mandatory from human studies with high evidence .The results of the present study suggest that the risk of EIF may be influenced by numerous factors. Additional research is required to better understand the mechanisms underlying these associations and to identify strategies for reducing the risk of EIF. This may entail replicating the study in a different sample, examining the role of other patient characteristics and treatment factors, and investigating the mechanisms underlying the associations between medical conditions and implant failure.Within the limitations of the present study, the use of AC is significantly associated with an increased likelihood of EIF. The OR was 2.64. Future research is required to validate and examine the prospective impact of AC on the osseointegration phenomena.


In this case, the filtration efficiency decreased by 11.8%, 7.98%, and 7.17%, respectively. The maximum difference in filtration efficiency of 0.29 μm particulates was about 12.7%. Eliminating static electricity slightly increased the resistance (2.5~15.5 Pa). In addition, the new reduced graphene oxide air filter material exhibited good conductivity and stability after continuous testing. This study provides data support for the application of subsequent electrification sterilization, reference values for multi-angle applications, and the development of new composite air filter materials.Improving the filtration efficiency of air filter materials is an ongoing research goal. This study conducted in-depth research on a new reduced graphene oxide air filter material, and the differences in its performance and conductivity durability before and after eliminating static electricity were tested and analyzed. The results showed that the filtration efficiency of the reduced graphene oxide air filter material significantly decreased after eliminating static electricity. The maximum decrease in filtration efficiency was observed at a filtration velocity of 0.8 m/s, with PM Complex air pollution has always been a key issue of concern ,2,3, as People are constantly trying various methods and developing new equipment to achieve this goal in response to the above demand. Air filters are widely used because they not only provide clean indoor air after treating outdoor harmful pollutants , but areAt present, many studies have been conducted on air filters ,10,11,12Ordinary air fiber filter materials mainly rely on mechanical blocking effects such as diffusion effect, interception effect, and inertia effect to filter particles in the air. As a result, the filtration efficiency is not significant for small particle sizes. For most bacteria and viral microorganisms, the size is on the micron or submicron scale. For example, relevant studies have shown that viruses are about 100 nm in size . These sTherefore, electret materials ,18 are cTherefore, this study focused on the above practical issues and conducted in-depth research on the differences in the performance and conductivity durability of a new reduced graphene oxide filter material before and after eliminating static electricity. It provides reference values for multi-angle applications and the development of new composite filter materials.Two clean, reduced graphene oxide air filter materials were selected as experimental materials in order to make the experimental purpose clearer and the experimental results more accurate; pieces with dimensions of 5 cm × 5 cm were tested. One piece was the blank group, where the static electricity was eliminated from the reduced graphene oxide filter material. Another piece was completely immersed in isopropanol for 2 min to remove the static electricity on the surface of the fiber . Next, i3, and the repeatability was 5%. An HD2114P.0 Portable Micromanometer was used to measure filtration resistance, and it was supplied by DeltaOHM Co., Ltd., Padua, Italy, with an accuracy of ±2% reading +0.1 m/s. The pressure range was ±0.4% F.S. An HD37AB1347 Indoor Air Quality Monitor was used to measure velocity, and it was supplied by DeltaOHM Co., Ltd., Padua, Italy, with an accuracy range of ±3%. A JSM-6510LV scanning electron microscope was used for analysis, and it was supplied by Japan Electronics Co., Ltd., Tokyo, Japan, its magnification was 5~30 million times, and its resolution was up to 3.0 nm. A TSI7525 Indoor Air Quality Meter Measuring Instrument was used to measure temperature and humidity, and it was supplied by TSI Instrument Beijing Co., Ltd., Beijing, China. The temperature measurement range was 0~60 °C, with a measurement accuracy of ±0.6 °C and 0.1 °C resolution. The relative humidity measurement range was 5~95% RH, measurement accuracy was ±3% RH, and the resolution was 0.1% RH. The average concentrations over 5 min before and after the testing were used in the calculations to reduce experimental errors [An experimental setup was built according to the experimental requirements , as showl errors .C1 is the concentration of particulate matter before filtration (μg/m3); and C2 is the concentration of particulate matter after filtration (μg/m3).The air filters’ filtration efficiency was calculated using Equation (1) .(1)η=C1P1 is the static pressure before filtration (Pa) and P2 is the static pressure after filtration (Pa).The filtration velocity before and after the filters were applied were the same, and the cross-sectional area was equal. The filtration resistance could be expressed by the static pressure difference. The filtration resistance was calculated using Equation (2) .(2)ΔP=PThe particle size distributions using outdoor atmospheric air as the dust source in this paper are shown in As shown in The filtration velocity range of 0.2~1.0 m/s was selected according to China’s national standard . The cha10 before eliminating static electricity was 55.3% to 63.1%, the filtration range for PM2.5 was 40.0% to 48.3%, and the filtration range for PM1.0 was 31.5% to 41.7%. Meanwhile, after eliminating static electricity, the filtration efficiency range for PM10 was 46.8% to 51.3%, the filtration range for PM2.5 was 35.7% to 41.1%, and the filtration range for PM1.0 was 27.9% to 33.7%. Thus, the filtration efficiency of the reduced graphene oxide air filter material after eliminating static electricity was significantly decreased compared to that before eliminating static electricity. The filtration efficiency for PM10 decreased by 8.5% to 11.8%, the filtration efficiency for PM2.5 decreased by 4.3% to 7.2%, and the filtration efficiency for PM1.0 decreased by 3.6% to 8.0%, with the greatest impact on PM10. This is because the isopropanol solution neutralizes the positive and negative charges on the fibers, causing their charges to permanently decay [10, PM2.5, and PM1.0 before eliminating static electricity was 63.1%, 48.3%, and 41.7%, respectively. After eliminating static electricity, the filtration efficiency of the new reduced graphene oxide filter material for PM10, PM2.5, and PM1.0 was 51.3%, 41.1%, and 33.7%, respectively. The filtration efficiency range of the reduced graphene oxide air filter material for PMly decay . After e10 > PM1.0 > PM2.5 . The main reason for this is that, after eliminating static electricity, the charges carried on the fibers are neutralized, resulting in a decrease in surface charge. This indicates that the filtration of fine particles mainly relies on electrostatic effects, while the filtration of large particles relies on other filtration mechanisms such as inertial collision. This conclusion is consistent with the literature [From terature  and indiThe counting filtration efficiency of the reduced graphene oxide air filter material before and after eliminating static electricity at the optimal filtration velocity is shown in The difference in filtration resistance of the reduced graphene oxide air filter material before and after eliminating static electricity is shown in From the figure, it can be seen that the filtration resistance of the reduced graphene oxide air filter material increased with the increase in filtration velocity before and after eliminating static electricity. Within the tested filtration velocity range, the resistance range before eliminating static electricity was 76~246 Pa, while the resistance range after eliminating static electricity was 78.5–261.5 Pa. Thus, the resistance of the reduced graphene oxide air filter material after eliminating static electricity was slightly higher than that before eliminating static electricity (a difference of 2.5~15.5 Pa). The main reason is that soaking the reduced graphene oxide air filter material in an isopropanol solution has a certain impact on the internal micropores. As a result, it causes a certain degree of contraction and binding between fibers, affecting the uniformity of the airflow velocity. The resistance cannot effectively pass through the fibers after colliding under the same air flow, resulting in an increase in its resistance, which is consistent with the conclusions in the literature . TherefoThe electric resistance characteristics of the reduced graphene oxide air filter material and its conductivity need to be investigated further. The fiber electric resistance values of the reduced graphene oxide air filter material and the variation in fiber electric resistance values over time are shown in From From the figure, it can be seen that an enveloping layer of reduced graphene oxide covered the surface of the reduced graphene oxide air filter material fibers, and some sections of reduced graphite oxide were cross-linked between the surfaces. This can provide a foundation for achieving fiber surface conductivity . In addiThe difference in the performance and conductivity durability of the new reduced graphene oxide air filter material before and after eliminating static electricity were measured and analyzed in this paper. The conclusions are as follows:10 decreased by 8.5% to 11.8%, the filtration efficiency for PM2.5 decreased by 4.3% to 7.2%, and the filtration efficiency for PM1.0 decreased by 3.6% to 8.0%, with the greatest impact for PM10. The degree of decrease was in the order of PM10 > PM1.0 > PM2.5 .The filtration efficiency of the reduced graphene oxide air filter material after eliminating static electricity significantly decreased compared to before eliminating static electricity. The filtration efficiency for PMThe counting filtration efficiency of the new reduced graphene oxide air filter material decreased significantly after eliminating static electricity. The filtration efficiency of 0~1.0 μm particles was the highest, with the efficiency ranging from 1.75% to 12.7%, while the difference was not significant for large particles. The maximum difference in filtration efficiency was for 0.29 μm particulates, at about 12.7%.The resistance after eliminating static electricity was slightly higher than that before (an increase of 2.5~15.5 Pa). As the number of days increased, it still exhibited relatively low electric resistance values and good conductivity.The electric resistance value after eliminating static electricity was greater than that before eliminating static electricity, with an overall increase of 7.0 KΩ to 9.4 KΩ and an average of 8.1 KΩ.


Biol. Lett.12: 20150935 (Published online 1 February 2016). (https://doi.org/10.1098/rsbl.2015.0935)Owing to a typographical error that was not caught in the page proofs, equation (3.3) is written incorrectly. The correct equation isc using the data in electronic supplementary material, table S1. Note that all angular values in the paper, including θ, are in degrees.This equation will recreate figure 2None of these errors affect the analyses or results presented in the paper.This has been corrected on the publisher's website.


Specificity and positive predictive values of DR for DN were 0.83 and 0.81, respectively . The Association of the duration of diabetes and the level of proteinuria with DN was statistically not significant (p > 0.05). Idiopathic MN (6) and Amyloidosis (2) were the most common isolated NDKD; whereas diffuse proliferative glomerulonephritis (DPGN) (7) was the commonest NDKD in mixed disease. Another common form of NDKD in mixed disease was Thrombotic Microangiopathy (2) and IgA nephropathy (2). NDKD was observed in 5(18.5%) cases in presence of DR. We noted biopsy-proven DN even in 14(35.9%) cases without DR, in 4(50%) cases with microalbuminuria and 14(38.9%) cases with a short duration of diabetes. Conclusion: Almost half (45%) of cases with atypical presentation have non-diabetic kidney disease (NDKD), though even among these cases with atypical presentation diabetic nephropathy  is commonly seen in 74.2% of cases. DN has been seen in a subset of cases without DR, with microalbuminuria, and with a short duration of diabetes. Clinical indicators were insensitive in distinguishing DN Vs NDKD. Hence, a kidney biopsy may be a potential tool for the accurate diagnosis of kidney disease.The heterogeneous spectrum of kidney disease in diabetes ranges from albuminuric or non-albuminuric diabetic kidney disease to non-diabetic kidney diseases. Presumptive clinical diagnosis of diabetic kidney disease may lead to an erroneous diagnosis. Material and Method: We analyzed the clinical profile and kidney biopsy of a total of 66 type 2 diabetes patients. Based on kidney histology, they were divided into—Class I (Diabetic Nephropathy), Class II (Non-diabetic kidney disease), and Class III (Mixed lesion). Demographic data, clinical presentation, and laboratory values were collected and analyzed. This study tried to examine the heterogeneity in kidney disease, its clinical indicator, and the role of kidney biopsy in the diagnosis of kidney disease in diabetes. Results: Class I consisted of 36(54.5%), class II 17(25.8%), and class III 13(19.7%) patients. The commonest clinical presentation was nephrotic syndrome 33(50%) followed by chronic kidney disease 16(24.4%) and asymptomatic urinary abnormality 8(12.1%). Diabetic retinopathy (DR) was present in 27(41%) cases. DR was significantly higher in the class I patients ( Traditionally, diabetic nephropathy (DN) is diagnosed clinically with evidence of proteinuria and diabetic retinopathy ,2,3,4. HThis retrospective cohort study included type 2 diabetes mellitus patients, who underwent kidney biopsies between October 2016 to October 2022. The study included a total of 66 cases, whose complete clinical data were available for analysis. Patient of age > 18 years and both gender  was included in the analysis. Patients of age < 18 years and with proteinuria < 30 mg per day were excluded from the analysis. All patients’ demographic data and clinical presentations were reviewed and recorded from hospital records. Fundoscopic evidence of diabetic retinopathy was noted for all patients. Laboratory value of urine analysis, 24 h urinary protein, Complete Blood Count, renal function test, liver function test, lipid profile, immunological marker , HBsAg, HCV, and HIV were obtained from records. Findings of light microscopy, immunofluorescence, and electron microscopy examination of kidney biopsy were reviewed and noted in detail for all patients. We analyzed the demographic data, clinical presentation, and laboratory value for the clinical syndrome, the indication of biopsy, and the type of nephropathy. Standard guidelines were used to define acute kidney injury (AKI), chronic kidney disease (CKD), and nephrotic syndrome ,12. Proteinuria was categorized into three categories: microalbuminuria (30–300 mg/day), Sub nephrotic (>300–3500 mg/day), and nephrotic range (>3500 mg/day) proteinuria. Based on kidney histology, patients were divided into three classes—Class I (Diabetic Nephropathy), Class II (Non-diabetic kidney disease), and Class III (Mixed lesion). Further analysis was done to find out the heterogeneity of kidney diseases, their clinical indicators, and the relevance of clinical indicators in the diagnosis of kidney disease in type 2 diabetes mellitus patients.p values to know the association between the clinical indicators and kidney histology. We calculated the sensitivity, specificity, positive predictive value, and negative predictive value of the clinical indicators for DN. Observation considered statistically significant for p-value less than 0.05.Statistical analysis was performed using PSPP version 1.4.1 . Descriptive statistics were presented as the mean and SD for continuous, and number and percent for categorical variables. A one-way ANOVA variance analysis test was used to examine the significance of the difference between the three classes. Pearson’s chi-square test was used to compare the three classes for categorical variables. We used multinomial logistic regression to estimate odds ratios (ORs), associated 95% confidence intervals (95% CIs), and p > 0.05) (p < 0.05). Isolated DN was seen in the majority of 22(81.5%) patients with diabetic retinopathy. However, NDKD either alone or in mixed form was noted in the remaining 5(18.5%) cases even in presence of DR. Isolated DN was noted in 14(35.9%) patients without DR. DN was the predominant lesion in presence of DR (81.5%), while NDKD either alone or in mixed form was a predominant lesion in the majority (64%) in the absence of DR. Sensitivity, specificity, negative and positive predictive value of DR for diabetic nephropathy are mentioned in the A total of 66 patients  with male to female ratio of 3.7:1 was included. The mean age of patients was 51.1 ± 10.5 years. The mean serum creatinine was 2.7 ± 1.8 mg/dL. The average proteinuria of 63 patients was 3.8 ± 2.8 gm per day. Three patients were anuric. The mean duration of diabetes was 7.1 ± 3.9 years . Based o > 0.05) . The pre > 0.05) . Diabeti > 0.05) . Idiopat > 0.05) . Of 8 cap > 0.05). Our findings regarding mean age, duration of diabetes, and 24 h urine protein excretions were like other studies [p = 0.175) and microalbuminuria  had a high risk of NDKD, but it was not statistically significant. Hence, recent data do not support the classical paradigm of diabetic kidney disease. Thus, our observation reasonably suggests that the level of proteinuria does not discriminate between DN and NDKD, and proteinuria is a poor predictor of the type of nephropathy in type 2 diabetes. The prevalence of diabetic retinopathy vary widely and ranges from 30–60% [p < 0.05). Isolated DN was seen in the majority of 22(81.5%) patients with diabetic retinopathy. Although, evidence of DR strongly suggest diagnosis of DKD but does not exclude the possibility of NDKD. Our findings also support the afore statement, because we noted biopsy proven NDKD  in the remaining 5(18.5%) cases even in presence of DR. Isolated DN was noted in 14(35.9%) patients in absence of DR, while the remaining 25 patients had isolated NDKD in 15(38.5%) and NDKD mixed with DN in 10(25.6%). DN was the predominant lesion in presence of DR (81.5%), while NDKD either alone or in mixed form was a predominant lesion in the majority (64%) in the absence of DR. Although, specificity and positive predictive values of DR for DN were high ; it had low sensitivity (0.61) and negative predictive values (0.64). We demonstrated that the absence of diabetic retinopathy was strongly associated with the presence of NDKD . Our finding also backs the observation of other published studies [In the present study, we retrospectively reviewed the demographic data, clinical features, and kidney biopsy of a total of 66 type 2 diabetes patients. The number of participants in the present analysis is low, which could be a reflection of a lack of awareness, resource limitation, limited access to a tertiary health care facility, and a reluctant strategy for a kidney biopsy at this center. This limits the power of the study and restricts the generalization of results. We found the mean age of the patient was 51.1 ± 10.5 years, and the female patient was 1421%). The mean duration of diabetes in our cohort was 7.1 ± 3.9 years. The average amount of proteinuria was 3.8 ± 2.8 gm per day. The mean serum creatinine was 2.7 ± 1.8 mg/dL. The mean age and mean duration of diabetes in Class II (isolated NDKD) were comparatively lower than Class I (DN) and Class III (DN + NDKD). However, the level of 24-h proteinuria in Class II (4.2 ± 3.3 gm) was comparatively higher than in Class I (4.0 ± 2.3 gm) and Class III (2.8 ± 3.2 gm). The mean serum creatinine in Class III was higher than in class I and Class II (3.8 vs. 2.4 vs. 2.6 mg/dL), reflecting either advanced or acute worsening of the disease. The three classes did not have statistically significant differences concerning age, sex, serum creatinine, proteinuria, and duration of diabetes  of cases with atypical presentation have non-diabetic kidney disease (NDKD), though even among these cases with atypical presentation diabetic nephropathy  is commonly seen in 74.2% of cases. About one-third (36%) of cases without DR have diabetic nephropathy, and about 20% of cases with DR have non-diabetic kidney disease on kidney biopsy. An almost equal number of cases have Diabetic nephropathy and NDKD in the presence of short diabetes duration. Factors like duration of diabetes, level of proteinuria, and presence of DR are not sensitive indicators for distinguishing DN vs. NDKD. Thus, clinical diagnosis alone may give an erroneous diagnosis. Hence, a kidney biopsy may be a potential tool for the early and accurate diagnosis of heterogeneous kidney disease in diabetes mellitus.The major limitation of this study was the small number of participants, participants with symptomatic disease, and Kidney biopsy in advanced disease.


Arthrobacter sp. strain ATA002 was isolated from seeds of the endemic cactus Maihueniopsis domeykoensis collected in the Atacama Desert, Chile. Here, we present a circular genome sequence, obtained by Nanopore sequencing, with a size of 3,904,590 bp and a GC content of 65.9%.The Gram-positive diazotrophic seed endophytic bacterium  The Atacama Desert is one of the most extreme environments on Earth . It is cMaihueniopsis domeykoensis were collected by AELA SpA and supplied by the Forestry Institute - INFOR  at Domeyko . The surface of the seeds was sterilized by sequential washing in 50% ethanol for 1 min, 2% sodium hypochlorite for 3 min, and 50% ethanol for 30 s, rinsing the seeds twice with sterile distilled water, grinding them in a clean mortar, and suspending them in combined carbon or Rennie semi-solid N-free medium . The DNA was cleaned using AMPure XP beads . The library was sequenced using the MinION platform and the Flongle flow cell (ONT). The sequencing ran for 72 h, and the quality was monitored using the MinKNOW v22.05.5 interface (ONT). Default parameters were used for all software unless otherwise specified. The raw sequencing data were base called and demultiplexed with super high accuracy using Guppy v6.0.6+8a98bbcbd (ONT), resulting in 100,206 reads with an Arthrobacter. The closest genome was Arthrobacter gandavensis strain JCM 13316 . A full-length 16S rRNA sequence comparison using NCBI BLAST (Arthrobacter koreensis strain DL (CP106856.1), with a query coverage of 99%. The genome was annotated using the NCBI PGAP v6.3 pipeline (The resulting genome was a circular contig 3,904,590 bp long, with a GC content of 65.9% and a coverage of 59×. The genome was found to be 94.15% complete and 0.98% contaminated. The taxonomic affiliation and closest related species were inferred using GTDB-Tk v2.1.0 (BI BLAST  revealedpipeline .Arthrobacter sp. ATA002 was deposited at GenBank under the accession number CP114015 and the BioProject accession number PRJNA894817. The raw reads are accessible via the SRA accession number SRR22099512.The genome of


Objective: Mycosis fungoides (MF) is the most common type of cutaneous lymphoma. The early stage of MF is a difficult diagnostic case, as it is often confused with many benign inflammatory dermatoses (BID). The study aimed to evaluate the diagnostic utility of TOX, FOXP3, CDD4 and GATA3 in differentiating early stages of MF from histologically overlapping BID lesions.Material and Method: A retrospective cross-sectional study was performed, in which immunohistochemistry (IHC) was used to evaluate the expression of TOX, FOXP3, CD4 and GATA3 in formalin-fixed paraffin-embedded (FFPE) sections of skin lesions from 30 cases with BID and 30 patients with early-stage MF.Results: The association between TOX expression and early-stage MF was statistically significant (P <0.001). TOX had the highest sensitivity of 96.77% and accuracy of 85.71% in diagnosis of MF; followed by CD4 with sensitivity of 85.71% and accuracy of 78.95%; and then, GATA3 with sensitivity of 76.7% and finally FOXP3 with sensitivity of 70.0%.Conclusion: TOX is suggested to be of higher diagnostic value in the early stages of MF than the conventionally used CD4 and other markers examined. Mycosis fungoides (MF) is considered the most common type of primary cutaneous T-cell lymphoma (CTCL) . It is vClinically, the disease progresses slowly in three stages including patches, plaques, and tumors . MF is hTranscription factor FOXP3 is used to identify normal regulatory T cells (Tregs) that have been assigned a variety of roles in immune regulation. An increase in normal FOXP3+ Treg cells in dermal infiltrates of MF has been linked to improved prognosis, similar to other indolent lymphomas . These cFOXP3 Tregs are found in various lymphomas as well as tumors . In a reGATA-binding protein 3 (GATA3), a transcription factor expressed in the epidermis, is essential for the regulation of epidermal differentiation and development of lymphoid cells; GATA3 is required in T helper (Th) cell subset differentiation into Th2 cells . GATA3 iThymocyte selection-associated high mobility group box factor protein (TOX), which belongs to DNA-binding factors, is involved in the formation of T cells. TOX is upregulated in the thymus during positive selection of CD4+ CD8+ precursors to CD4+ T cells, but it shows a decreased expression in mature CD4+ T cells after leaving the thymus . The difTherefore, in this work we aimed to evaluate the diagnostic utility of common immunohistochemical markers, including TOX, FOXP3, CD4 and GATA3, used in diagnosing early-stage MF in order to avoid misdiagnosis and thus ensure early proper management.This retrospective cross-sectional study included 30 cases of early MF and 30 cases of BID  collected from the archives of the Department of Pathology, Zagazig Faculty of Medicine (2018-2021).Patients’ consent was obtained; and the research was reviewed and approved by the Institutional Research Board (IRB), (ZU-IRB #6967/-2-6-2021). The research complied with the Helsinki Declaration.MF was diagnosed by looking at the histologic features of sections stained with hematoxylin and eosin (H&E). All of the included cases of MF had an active lesion in the early stage; and BID cases were also with an active lesion. Pregnant and lactating women and patients with the tumor stage of MF or other types of cutaneous B or T-cell lymphomas were excluded.The diagnostic feature of MF was the predominance of atypical lymphocytes whose nuclei are larger than those of dermal lymphocytes. The atypical cells had an irregular nuclear border surrounded by a clear halo and were observed in the basal layer or in both stratum basalis and stratum spinosum layers as single cells. Pautrier’s microabscesses were also detected. Clinical findings included the impressions of dermatologists, and the IHC findings (pan T cell markers) were present in the reports.Analysis for TOX, Fox and GATA3 was carried out on 5-μm thick archived formalin-fixed paraffin-embedded (FFPE) sections using the Leica Bond III autostainer. The primary antibodies were applied to the slides. They included Rabbit polyclonal anti-TOX antibody, as the optimal primary antibody, at a dilution of 1:150  and GATA3 mouse monoclonal antibody , CD4 , CD8  and FOXP3 mAb . Appropriate negative and positive controls were included. The slides were examined by three pathologists separately.The IHC evaluation was done by reporting the number of cells with nuclear  or cytoplasmic-membranous (CD4) staining per 10 high power fields (HPF) in both the epidermis and dermis; and the positive cell percentage was evaluated as Grade 0 if no cells were stained, Grade 1 if < 10%, Grade 2 if 11–50% and Grade 3 if > 50% of cells stained ,14.Statistical analysis was assessed using SPSS software, version 11.5 . The Pearson χ2-test and Fisher exact test were used for comparison of the IHC results between the MF and nonmalignant groups. The p value of ≤0.05 was accepted as statistically significant. Sensitivity, specificity, false negative and positive rates (1−sensitivity), and diagnostic accuracy of each marker were assessed.Sixty cases (30 MF and 30 BID) were included in this study. Evaluation revealed higher grades (2 and 3) of TOX immunohistochemical marker expression in MF cases (33.4% and 60% respectively) than in BID, in which 70% of cases were negative for TOX expression. The results were highly significant (p=0.001) .GATA3 expression assessment revealed that negative and grade 3 GATA3 expression were higher in MF cases being 23.3% and 66.7%, respectively, compared with BID of 3.3% and 43.4%, respectively; and the results were highly significant (p=0.002). Meanwhile, Grades 1 and 2 GATA3 immunohistochemical expressions were significantly higher in BID (33.3% and 20% respectively) than found in MF cases (6.7% and 3.3%), and the results were highly significant (p=0.002) .Regarding assessment of CD4 expression, we found that 40% of MF cases showed grade 3 expression while among the BID cases only 6.67% showed it, and the results were significant (p=0.0003). However, negative CD4 immunohistochemical expression was significantly higher in BID (63.3%) than in MF cases (16.7%) .Regarding FOXP3 expression, we found that grades 1 and 3 of FOXP3 expression were higher in MF cases  than in BID cases (23.3% and 20% respectively), and the results were highly significant (p=0.002). However, negative and grade 2 FOXP3 expressions were significantly higher in BID  than in MF cases (13.3% and 0.0% respectively); and the results were highly significant (p=0.002) .The immunohistochemical assessment revealed higher expression (grade 3) of TOX, GATA3, CD4 and FOXP3 in MF than in BID. Negative expressions of TOX, CD4 and FOXP3 were significantly higher in BID than MF cases, but the reverse was true for GATA3 negative expression as it was significantly higher in MF than BID. Moreover, significant differences in other grades between MF and BID were detected (p<0.05) .  followed by CD4 (85.71%), GATA3 (76.7%) and finally FOXP3 (70.0%). Regarding the specificity, TOX had the highest specificity (76.92%) followed by CD4 (73.17%), FOXP3 (13.3%) and finally GATA3 (3.3 %). The marker with highest positive predictive value (PPV) was TOX with 76.92% followed by CD4 (73.17%), FOXP3 (44.7%) and finally GATA3 (44.2%). The marker with highest negative predictive value (NPV) was TOX 96.77% followed by CD4 (85.71%), FOXP3 (30.8%) and finally GATA3 (12.5%). Among the studied markers, TOX had the highest diagnostic accuracy (85.71%) followed by CD4 (78.95%), FOXP3 (41.7%) and GATA3 (40.0%). To sum up, TOX had the highest sensitivity, specificity, PPV, NPV and diagnostic accuracy followed by CD4. FOXP3 was following CD4 as regards specificity, PPV, NPV and diagnostic accuracy but it was less sensitive than GATA3 in distinguishing MF from BID. In other words, GATA3 had the lowest specificity, PPV, NPV and diagnostic accuracy but it was more sensitive than FOXP3 in MF diagnosis than BID .The early stage of MF is a challenging clinical condition, as it is often confused with BID. The difficulty in differential diagnosis between early MF and BID may be due in part to a lack of tumor cell-specific markers . FindingTOX is one of the proteins that act as a gene expression regulator by changing the structure of chromatin . RegardiThe FOXP3 gene encodes a transcriptional factor that is vital for the function and development of regulatory Tregs, which are differentiated from naive CD4+ T-helper cells . MutatioIn line with previous reports, the current work revealed that the difference between MF and BID regarding FOXP3 expression was statistically significant . Wada etThe function of FOXP3 in MF development and progression is quite puzzling. Manso et al.  reportedExpression of GATA3 showed a significant difference between MF and BID. This can be explained by the fact that TH1 and not TH2 is the prominent intraepidermal cell in early MF, and GATA3 stains the later one .According to our results, TOX is recommended as a marker for diagnosis of early-stage MF, helping to differentiate it from the benign mimickers. This is because TOX had a high sensitivity (96.77%) and accuracy (85.71%), followed by CD4 for sensitivity (85.71%) and accuracy (78.95%). The same results were obtained by Zhang et al. . MorimurThe limitations of the study included the relatively small numbers of cases seen at a single medical center. Furthermore, the study did not include all types of BIDs.Among the immunohistochemical markers examined, TOX is suggested to be the most accurate in diagnosing MF, which is difficult to diagnose especially in its early stages when it is confused with BID. Further future studies including larger numbers of patients are recommended to confirm the diagnostic significance of TOX.The authors declare that there is no conflict of interest regarding the publication of this paper.No funding has been received.


The COVID-19 pandemic has been linked to chronic pulmonary complications all over the world. Respiratory complications such as chronic cough, dyspnea, increased respiratory rate, and oxygen support demand are prevalent in recovered COVID-19 patients. These problems are long-term and have a negative impact on one’s quality of life. Patients must be evaluated for potential complications, and risk factors must be found. Some reports around the world explain the factors that contribute to the development of these complications. However, to the best of our understanding, no reports of post-COVID-19 complications have been reported from Ethiopia.Facility based cross-sectional study was done among 405 participants selected by simple random sampling technique. Structured questionnaire which includes participants’ demographic, clinical and 3rd month visit characteristics was collected by Open Data Kit and exported to SPSS version 25.0 for analysis. Percentage with frequency and median with Interquartile range was used in descriptive statistics. The association between variables was analyzed with bivariate and multi variable logistic regression. A statistical significance was declared at p-value < 0.05, with 95% confidence interval.The median (Interquartile range) age of participants was 57.0  years, 63.2% were males. The prevalence of post-COVID-19 pulmonary complication in recovered COVID-19 patients was 14.1% . After adjusting for possible confounders on multivariate analysis, older age  and consolidation  were shown to have significant association with post COVID-19 pulmonary complications.The prevalence of post COVID-19 pulmonary complication was observed to be lower than other reports globally. Older age and the presence of consolidation on lung imaging were associated with those complications. Clinicians are recommended to consider assessing the lasting effects of the pandemic, beyond immediate care, and should also investigate the COVID-19 history in patients presenting with respiratory issues. The COVID-19 pandemic is observed to have long-lasting impacts on the respiratory system, which is primarily affected in the acute phase of infection. In recovering patients, it is common to observe a cough, shortness of breath, decreased respiratory functions, a need for oxygen support, and radiologic evidence of chronic lung inflammation .The care and follow-up of infected patients should continue after the acute phase, as an important proportion of patients continue to have persistent symptoms and complications, as can be learned from the previous coronavirus-originating epidemics –7. GivenPost COVID-19 pulmonary complications have been reported around the globe. Multiple reports from US, Italy and other countries suggest that the prevalence of these symptoms is high, some reporting as high as 87.4% –14. FatiFacility based cross sectional study design was conducted to assess the prevalence and associated factors of post COVID-19 pulmonary complications among recovered COVID-19 patients in Eka Kotebe General Hospital, which is the largest treatment centre in the country. The hospital had post-COVID follow up clinic which is run by general practitioners, consultant internists and a pulmonologist. The data was taken from the medical records of patients who, between January 2021 and January 2022, had their third month follow-up appointment after being admitted with confirmed COVID-19 and were discharged as recovered.The sample size was calculated by using single population formula with the assumptions of: 95%CI, 5% margin of error, with 40.2% expected proportion . 10% wasThe data was collected by using pretested structured checklist which is adopted from reviewing different related literatures , 17, 19.Presence of 2 or more of persistent cough, persistent shortness of breath, increased respiratory rate (> 22 breaths per minute) and requirement of oxygen support (SpO2 < 90% on room air) was used to define post-COVID-19 pulmonary complication during 3rd month visit , 13, 19.Data was collected using ODK software and, transported to and analyzed using SPSS software version 25.0. Categorical variables were presented as frequency and percentages (%), and continuous variables were described using the median and interquartile range, after checking for skewness. The association of independent variables with the outcome variable was investigated using binary logistic regression analysis. The variables that showed an association with the outcome variable at the bi-variable analysis with p value < 0.25 were entered into the final multivariable analysis to control for potential confounders. Adjusted odds ratio (AOR) along with 95% confidence interval was estimated to assess the strength of association and a P value < 0.05 was considered to declare the statistical significance in the multivariable analysis. Assumptions like model fitness was checked to be satisfied using Hosmer and lemeshow p-value .A total of 405 study participants were involved, 256 (63.2%) were males. The median (IQR) age of the participants was 57.0  years Table .Majority of the participants were symptomatic during admission. Cough (86.9%), easy fatigability (69.4%), loss of appetite (54.8%), and fever 53.1%) were the most common symptoms presentation at admission. Shortness of breath and headache were also recorded in 46.2% and 34.6% of participants respectively  was done for 322 (79.5%) participants. The most common findings were ground glass opacities (GGO) and consolidation. Consolidation was found in 159 (39.3%) of participants, while GGO was present in 274 (67.7%). Infiltration and effusion were also found in 69 (17.0%) and 7 (1.7%) participants respectively (Table The median (IQR) hemoglobin level was 14.6 g/dl 13.3, 15.8) and the WBC count was 7530  cells per microliter. More than half (51.4%) of participants had a low lymphocyte count (ALC) of less than 1000 per microliter. The median (IQR) lymphocyte count was 973  cells per microliter. The median (IQR) creatinine and BUN level were 0.91  mg/dl and 14.0  mg/dl respectively  had severe COVID-19 diagnosis during their hospital stay. Twenty eight (6.9%) had critical COVID-19, 29 (7.2%) had moderate COVID-19 and 21 (5.2%) had mild COVID-19. Majority (87.7%) were admitted to the medical ward while 8.9% of participants admitted to ICU/HDU. The remaining 14 (3.5%) participants were admitted to other wards like the labor ward or psychiatry ward.Participants received different levels of oxygen support during their stay. Forty seven (11.6%) participants were maintaining their oxygen saturation with room air, without any support, 70.4% of them were supported with 1-5 L per minute of oxygen, 47 (11.6%) participants were given 6-15 L per minute while 5.2% (n = 21) were put on non-invasive ventilation (NIV) support. Only 5 (1.2%) of participants were put on a mechanical ventilator (MV). Steroid was given to 92.1% (n = 373) of participants and 86 (21.2%) participants received antiviral treatment for COVID-19.The median (IQR) length of stay was 11 , 15 daysThe median (IQR) number of days elapsed from the date of discharge to the visit day was 88  days. During this visit, 72 (17.8%) participants had respiratory complaints. 57(14.1%) had shortness of breath and 31(7.7%) had a cough. Among these symptomatic patients, 16 (4%) participants complained of both cough and shortness of breath, while 56 (13.8%) of them had only one respiratory complaint.The respiratory rate of participants during the 3rd month visit ranged from 10 to 38 breaths per minute, with a mean number of 22.29 (SD = 2.87), and participants with a respiratory rate of more than 22 breaths per minute were 42.2% (n = 171) of all participants. Oxygen saturation measured at room air during the visit also ranged from 72 − 99%, with a mean measure of 95.54% (SD = 2.74). 23 (5.7%) participants had an oxygen saturation level of less than 90% , 135 (33.3%) of participants had one or more of these complications. Two of the complications were present in 7.7% (n = 31) of participants, while 19 (4.7%) had three complications. 1.7% (n = 7) of participants had all the above four complications. The proportion of participants with 2 or more pulmonary complications which is used to define the prevalence of post COVID-19 pulmonary complications is 14.1% (n = 57) with 95% CI: 10.8%, 17.8%.New onset comorbid illnesses, which were not diagnosed during admission were also reported during this period, the most common being DM and hypertension. They were reported in 11.6% (n = 47) and 9.4% (n = 38) of participants respectively. Cardiac disease was found in 10 2.5%) and CKD was diagnosed in 8(2.0%). GGO was found in 19 (4.7%) and consolidation was seen in 3(0.7%) of participants (Table % and CKDThe median (IQR) WBC count during 3rd month visit was 6530  cells per microliter. Participants with lymphocyte count less than 1000 cells per microliter were 7.9% (n = 32). The median (IQR) lymphocyte count was 1776  cells per microliter .Participants whose age is less than 40 years are 77.3% less likely to develop post-COVID-19 pulmonary complications than those aged greater than 60 years . There is also a 59.9% reduced odds of developing complications in participants aged from 41 to 60 years . Participants whose chest imaging did not show consolidation were 50.3% less likely to develop pulmonary complications after discharge than their counterparts [AOR = 0.497, 95% CI (0.258–0.957)  cut point used to define long- COVID-19 in most literatures. These literatures showed a higher prevalence of post COVID-19 complications, up to 39% prevalence report . In our Shortness of breath on the day of visit was observed in 14.1% participants, which was close to the Egypt study that reported 17.4% . It is mCompared to majority of reports, this study has a lower prevalence rate of pulmonary complications. A meta-analysis on long COVID by A.V.Raveendran et al. reported around 35% prevalence of residual symptoms at 3 months follow up . A 32% pNew onset comorbidities, which were not recorded during admission, were observed in participants during the visit, DM and hypertension being the commonest ones with 11.6% and 9.4% prevalence. Cardiac illnesses were observed in 2.5% participants. This is comparable to other studies which reported new onset comorbid illnesses after COVID infection. A systematic review and meta-analysis on new onset diabetes reported 19.7% new-onset DM and 25.23% hyperglycemia prevalence .Regarding the factors associated with development of pulmonary complications, age greater than 60 years was observed to be at more risk than those lower than that age. There was a 77.3% less chance of pulmonary complications in participants whose age is less than 40 years and 59.8% reduced odd of developing complications in participants aged from 41 to 60 years. This finding was similar to some studies globally, which reported older age as a risk factor for pulmonary complications , 22, 25.symptoms . This caAmong the admission status of participants, presence of consolidation was the one factor that showed significant association with post-COVID-19 pulmonary complication, participants with no consolidation in their chest imaging being protected by 50.3%. Extensive chest imaging finding were observed to be associated with pulmonary complications in other studies too . PatientThe prevalence of post-COVID-19 pulmonary complication in recovered COVID-19 individuals is lower than reported elsewhere. Age > 60 years and consolidation in chest imaging have a significant association with pulmonary complications. Clinicians must evaluate the long-term impact of the pandemic, in addition to acute management, and look for a history of COVID-19 infection in patients with pulmonary complaints.


Aim: The national Olympic committee of Switzerland has conducted an online survey among female elite athletes with a focus on cycle disorders, contraception, and injuries in 2021.Methods: A total of 1,092 female elite athletes from 107 different sports were asked to answer the questionnaire. A descriptive analysis was carried out to determine location parameters and create frequency tables.Results: The questionnaire was completed by 408 athletes (37.4%) from 92 different sports. 43.4% participated in a lean sport. 57.1% reported no injuries, 32.6% one injury, and 10.2% two or more injuries per year. A considerable proportion reported being affected by primary amenorrhoea (10.8%). Primary amenorrhoea occurred significantly more often in female athletes with a BMI lower than 21.7 kg/m2 (15.2%) than in athletes with a BMI above 21.7 kg/m2 . Considering contraception, 25.8% of female athletes were currently using an oral contraceptive pill. The proportion of female athletes not using contraception at all or using non-hormonal contraceptive methods was high at 54.4%. In lean sports, significantly more athletes used no or non-hormonal contraceptives (p < 0.05).Conclusion: Among top Swiss female athletes, a considerable proportion used non-hormonal or no contraceptives. This trend was more evident in lean sports. Delayed menarche and cycle irregularities were common among female athletes, especially among athletes with high training volumes as well as a BMI below 21.7 kg/m2. This orienting survey underlines the importance of specialized gynecological care for elite female athletes. In so-called lean sports, performance is linked to body weight and body composition. This is reflected in a higher rate of cycle disorders (up to 79%) in these sports . Lean spversus non-lean sports was done separately by the authors. The respective classification was then compared in a four-eye procedure and a consensus was found  general information, 2) injuries, 3) menstrual cycle, 4) urinary incontinence, 5) contraception, 6) pregnancy, 7) nutrition, and 8) taboo topics. A total of 1,092 female elite athletes from 107 sports were emailed and invited to participate in the study. Of these, a total of 408 female elite athletes completed the online questionnaire. The questions on contraception were answered by 401 of 408 female elite athletes. The questions on menarche were answered by 388 of 408 female elite athletes with an age statement. The question on secondary amenorrhea was answered by 405 of 408 female elite athletes. All female elite athletes answered the questions about injuries.versus non-lean sports, individual versus team sports) differed in the endpoints tested, Pearson’s chi-squared test was used. The Mann-Whitney U test was used to test group differences and distributions for metric measurement data. The effects of multiple testing were not considered in the descriptive analysis. Statistical significance was defined as p-value <0.05.Frequency data were used to analyse ordinal and nominal scaled variables descriptively. Mean (standard deviation) and median (quartile 1/quartile 3) were determined for metric data. To examine whether subgroups  had to have been answered. Answers like “I do not know” were considered as unspecified data. Half of the female elite athletes were 23 years old or younger, 30% were 24–29 years old and 20% were 30 or older. Concerning training volume, more than 60% of female elite athletes state, that they train 10–20 h weekly (n = 245), 10.8% less than 10 h weekly (n = 41), and 28.8% more than 20 h weekly (n = 112). In the survey, female elite athletes from individual sports  were more heavily represented than female elite athletes from team sports  . Since fversus non-lean sports and the contraceptives were divided into the following three groups: 1) non-hormonal and no contraception, 2) systemically hormonal contraception  and 3) hormonal intrauterine devices (LNG-IUD). The LNG-IUD was considered separately due to the low serum LNG concentrations (0.1–0.4 ng/mL) and the consequently negligible systemic effects  were significantly less (p = 0.049) likely to use systemically active hormonal contraceptives  than female athletes from non-lean sports . Female athletes in lean sports were more likely to prefer an LNG-IUD , but not statistically significant (p = 0.197) or to report the use of non-hormonal or no contraception. The 181 female athletes were asked about their reason for using hormonal contraception. The following multiple-choice answers were possible: 1) contraception, 2) reduction of dysmenorrhea, 3) cycle control, and 4) reduction of bleeding intensity. The most frequent reason given for hormonal contraception was the desire to use a contraceptive , followed by the possibility of reducing menstrual cramps , cycle control in competitions , and reduction of bleeding volume . Some female athletes mentioned treatment of acne , treatment of endometriosis , iron deficiency , regular bleeding pattern , low cost  and stabilisation of body weight  as further individual reasons for using hormonal contraception. The question of whether the hormonal contraceptive pill influences performance was answered neutrally to positively by most of the female athletes. In terms of numbers, 31.5% (n = 57) felt no difference, 43.6% (n = 79) could not answer the question (“I do not know”), 14.4% (n = 26) thought their performance was more consistent, 1.1% (n = 2) felt it was less consistent, 7.7% (n = 14) felt more powerful, and only 2.8% (n = 5) felt their performance was worse with hormonal contraceptive methods in general.Among the Swiss female elite athletes surveyed, 45.1% (n = 181 of 401) of the respondents used hormonal contraceptives and of those 57.5% (n = 104 of 181) used an oral contraceptive pill (OCP). OCP refers to both progestin-only pills (POP) and combined oral contraceptive pills (COC). Among the remaining female athletes with hormonal contraception, 9.4% used a vaginal (n = 17) ring, 32.0% (n = 58) used a levonorgestrel intrauterine device (LNG-IUD) and 1.1% (n = 2) used a progestin-only implant. For the contraceptive pill, no further differentiation was made between the combined contraceptive pill (COC) and progestin-only pill (POP). 54.9% (n = 220 of 401) of the study participants used no or a non-hormonal contraceptive method. In this survey, non-hormonal methods were not broken down further . For further analysis, the 92 sports were divided into lean sports  effects . Using tversus 9.2% . When comparing individual and team sports (p = 0.861). We performed the same calculation based on BMI and divided the athletes into two groups: the normal/overweight group  and the underweight group . In this further examination of menarche onset as a function of BMI , no statistically significant differences were found (p = 0.611) due to the small number of women with primary amenorrhoea in the underweight group (n = 10). To examine the function of BMI on menarche in even greater detail, new BMI subgroups were defined based on the median of BMI (21.7 kg/m2), i.e., female athletes with a low BMI  versus female athletes with a high BMI . Examination of the frequency of primary amenorrohea as a function of this BMI classification showed that primary amenorrhoea occurred significantly more often in female athletes with a lower BMI  than in athletes with a higher BMI .Menarche refers to the age at which menstrual bleeding first occurs. On average, this happens between the ages of 12 and 13 . Primarym sports , there wOligomenorrhoea is defined as fewer than nine cycles per year or a cycle length of more than 35 days in adults or more than 45 days in adolescents . Overallversus non-lean sports. We also examined the influence of training volume and secondary amenorrhoea on the frequency of injuries. A total of 398 athletes reported their training volume, and 387 athletes reported both their training volume and information about a current or past episode of secondary amenorrhoea.All female athletes surveyed answered the question about an injury in the past year. 57.1% (n = 233) of the respondents reported that they had not been injured, while 32.6% (n = 133) reported at least one injury and 10.2% (n = 42) reported two or more injuries per year. The risk of injury did not differ when comparing lean p = 0.004). In contrast, there were no statistically significant differences in women without secondary amenorrhoea regarding injuries at different training intensities . Accordingly, female athletes in lean sports with a current or past episode of secondary amenorrhoea and higher training volumes (≥21 h per week) had a significantly higher risk of injury.To investigate differences in the incidence of injuries in female athletes with secondary amenorrhoea, we divided the data of these 387 female athletes into two groups: 1) less than 21 h of training per week  and 2) at least 21 h of training per week . Female athletes affected by secondary amenorrhoea and with high training volumes (≥21 h/week) were significantly more likely to report at least one injury per year , whereas female athletes with a current or past episode of secondary amenorrhoea and lower training volumes (<21 h/week) tended to report fewer injuries . With 408 elite female athletes participating, the results reflect a representative sample of Swiss elite athletes. In the context of the informal request for study participation within a newsletter by email through Swiss Olympic, the response rate of 37% is be regarded as significant.Our online cross-sectional survey of 92 different sports in Swiss female elite athletes shows that the incidence of primary amenorrhoea (11%), secondary amenorrhoea (5.4%), and oligomenorrhoea (19.1%) in Swiss female elite athletes remains a problem compared to the incidence in the normal population  and alsop = 0.010) higher in lean sports (26.7%) than in non-lean sports (16.6%)  of Swiss female elite athletes. Overall, 11.3% of the Swiss female elite athletes surveyed stated, that their menstruation started late, i.e., after the age of 14, which corresponds to primary amenorrhoea . Accordi (16.6%) . Our res (16.6%) . Athlete (16.6%)  are at g (16.6%) . Delayed (16.6%) . 396 par (16.6%) . A surve (16.6%) . This wi (16.6%) . In comp (16.6%) . Prolong (16.6%) . In addi (16.6%) . Regularversus 43.2% in the general population . An OCP was used by 25.9% of the female athletes (COC or POP), 14.5% used an LNG-IUD, and 4.2% a vaginal ring. Compared with the normal population in Switzerland, it seems that OCPs are less popular among Swiss female elite athletes. According to the 5-yearly representative survey of 3,555 Swiss women (aged 15–49) in 2017, the condom was used most frequently (42%), followed by the OCP  . This trpulation  used OCPpulation , comparepulation . Similarpulation , Norway pulation , and the the USA . In the  the USA . The con the USA , 72% of  the USA , and 57%use OCPs , compareuse OCPs  and 14% use OCPs . These duse OCPs . Combineuse OCPs . Becauseuse OCPs . The infuse OCPs . In thisAll female athletes surveyed answered the question about an injury in the past year. 57.1% of the female athletes reported that they had not been injured, 32.6% reported at least one injury and 10.2% reported two or more injuries per year. Female athletes in lean sports with a current or past episode of secondary amenorrhoea who trained intensively (≥21 h per week) had a significantly higher reported incidence of injury. Other studies have also shown that female athletes in lean sports are at greater risk of injury . AccordiThe results of this orienting survey among Swiss female elite athletes show that contraceptive behavior in Swiss female elite athletes differs in comparison to the normal population. The majority of Swiss female elite athletes used no or non-hormonal contraceptive methods.The reason for different contraceptive choices is yet to be determined. According to the literature, a possible explanation for a declining rate of hormonal contraceptives could be a lack of information in both the women and the care-taking physicians concerning the needs of each individual patient . Further2) appears to be very low in this survey . If the proportion of underweight female athletes is underrepresented due to missing data, an even higher proportion of female athletes with primary or secondary amenorrhoea would be expected. When choosing a different BMI classification using the median (BMI <21.7 kg/m2versus BMI >21.7 kg/m2), a statistically significant accumulation of delayed menarche depending on BMI could be demonstrated (p = 0.021). This finding would have to be substantiated in larger samples. Another limitation of this study is the lack of assessment of energy deficiency among the Swiss female elite athletes surveyed. In the questions on OCPs, no further distinction was made between combined (COC) and progestin-only pills (POP) or other contraceptive methods such as, for example, the hormonal patch, for example,. Similarly, the non-hormonal contraceptive methods lacked a differentiation between the various non-hormonal methods . According to the literature, lean sports are associated with an increased risk of RED-S-related symptoms such as cycle disorders and injuries , a certain residual blurring of the distinction could not be completely avoided.The proportion of underweight female athletes (BMI <18.5 kg/minjuries . The aim


BackgroundAcute appendicitis is the most common surgical emergency worldwide with scarce reports about its prevalence in the Middle East. To date, no epidemiological article has described the incidence of appendicitis in Lebanon. Our primary objective was to estimate the rate of appendicitis in a single center in Lebanon. Our secondary objectives included identifying differences between simple and complicated appendicitis regarding demographics, pre and postoperative characteristics, and symptoms and signs of appendicitis.MethodologyA retrospective study was conducted at a single central university hospital in Lebanon. Patients with a clear diagnosis of acute appendicitis were included. Pregnant women, lactating women, patients with organ dysfunction, and patients younger than 18 years old or older than 80 years old were excluded. We reviewed and collected the data of patients who presented to the hospital between November 2018 and November 2019 and November 2020 and November 2021.Results2 compared to 18.97 ± 10.37 kg/m2 in patients with complicated appendicitis (p = 0.94). A total of 42.3% of patients who used antibiotics 24 hours after the operation had simple appendicitis, whereas 20.8% had complicated appendicitis (p = 0.004).A total of 95 patients were included in our study, with 35 women and 60 men. The mean body mass index of patients with simple appendicitis was 19.14 ± 9.66 kg/mConclusionsAntibiotic usage and the length of hospital stay were correlated with the severity of appendicitis, as reported in the literature. Further randomized studies with a larger number of patients and covering several hospitals in Lebanon are warranted. Acute appendicitis (AA) is known to be the most common surgical emergency worldwide . A lowerThe diagnosis of AA can be quite challenging. The 2020 update of the World Society of Emergency Surgery Jerusalem guidelines recommends the use of an individualized approach for the diagnosis of AA and for risk stratification based on sex, age, and symptoms . The manA recent population-based study estimated the incidence of appendicitis in the 21st century to be 100, 105, 150, and 160 per 100,000 individuals in North America, Eastern Europe, Western Europe, and Turkey, respectively . The stuWe believe that studies on the epidemiology of appendicitis are much needed as they will pave the way for future studies on the economic burden of appendicitis, related to appendectomy and hospital stay. The usage of fried and fatty meals in traditional Lebanese cuisine is well known for raising the risk of appendicitis. In addition, the identification of factors related to complicated appendicitis is the first step toward the implementation of strategies aimed at reducing the incidence of complications arising from AA.Study design and ethical approvalA retrospective review of patient charts was performed. Patients admitted with a diagnosis of AA in the period between November 2018 and November 2019 (before the COVID-19 pandemic) and between November 2020 and November 2021 (during the pandemic) were included. Patients were divided into two groups, namely, simple appendicitis and complicated appendicitis. All patients were treated at Al Zahraa University Medical Center in Beirut, Lebanon. Ethical approval was obtained from the Institutional Review Board at Al Zahraa Hospital University Medical Center, Beirut, Lebanon . Informed consent was not required because there was no risk to participants and the study only conducted a retrospective analysis of patient records.Inclusion and exclusion criteriaInclusion and exclusion criteria were similar to a previously published study by Bazzi et al. . LebanesGeneral characteristics2 versus 18.9 ± 10.3 kg/m2 for patients with complicated appendicitis . Sex, age, and BMI were statistically non-significant with respective p-values greater than 0.05 .A total of 95 patients were included in this study, with 35 females and 60 males. Patients with simple appendicitis had a mean age of 33.7 ± 13.3 years, whereas those with complicated appendicitis were older with a mean age of 39.1 ± 13.5 years (p = 0.08). The mean body mass index (BMI) of patients with simple appendicitis was 19.1 ± 9.6 kg/mThe mean C-reactive protein (CRP) level was 111.4 ± 34.1 mg/dL among patients presenting with simple appendicitis versus 150.3 ± 66.5 mg/dL among those having complicated appendicitis. This association was statistically significant with a p-value of 0.03 (<0.05) .Finally, right lower quadrant pain, migration/periumbilical pain, nausea, fever, and anorexia were the most commonly reported signs and symptoms of AA Table .Postoperative characteristicsFirst, among patients who received antibiotics for 24 hours postoperatively, 42.3% had simple AA and 20.8% had complicated AA. Second, among those who received antibiotics for 48 hours postoperatively, 46.5% had simple AA and 37.5% had complicated AA. Third, among those who received antibiotics for three days postoperatively, 8.5% had simple AA and 29.2% had complicated AA. Finally, those who received antibiotics for five to seven days postoperatively had complicated AA only. This association was statistically significant with a p-value of 0.004 (<0.05)  transit in one day, 42.3% had simple AA and 12.5% had complicated AA. Among those who had bowel movements in two days, 49.3% had simple AA and 70.8% had complicated AA. Additionally, among those who had GI transit in four days, 4.2% had complicated AA with none belonging to the simple appendicitis group. Moreover, the majority (41.7%) of those who had their abdominal abscess drained had complicated appendicitis primarily. These associations were statistically significant with a p-value of less than 0.05.The mean hospital stay was 2.69 ± 0.919 days in cases of simple AA compared to 3.88 ± 1.84 days in cases of complicated AA. This association was equally statistically significant with a p-value <0.01 . According to American guidelines, antibiotic use is advised for four to seven days after complicated appendicitis . PostopeMoreover, in our study, intraperitoneal cultures were taken more in cases of complicated appendicitis. This is very important to prevent uncommon infections that might lead to more severe symptoms, especially with the increase in drug resistance. Bacteria resisting the chosen antibiotics means more infectious complications and longer hospital stays among patients with appendicitis ,22. Our Furthermore, longer hospital stays were seen in cases of complicated appendicitis rather than in cases of simple appendicitis. This is in line with other studies reporting that patients with complicated AA stay six times longer in hospitals when compared to those with simple AA . SimilarIn our study, among those who underwent drainage for an appendiceal abscess, the majority had complicated appendicitis. This is consistent with previous studies stating that patients with complicated appendicitis and who underwent appendectomy are more prone to develop postoperative complications such as abscesses . This caOne of the main limitations of our study is collecting data from a single center. Thus, our data cannot be extrapolated to the country, even though it is one of the largest hospitals. Additionally, the COVID-19 pandemic and the retrospective nature of the study are also other limitations. A selection bias could be present due to the loss of follow-up with some of our included patients, and missing data in patient files could have predisposed to information bias.CRP levels, antibiotic usage, and length of hospital stay were correlated with the severity of appendicitis, which is similar to other studies reported in the literature. Further randomized studies with a larger number of patients and covering many hospitals in Lebanon are warranted.


Single- and dual-species biofilms of Enterococcus faecalis and Candida albicans were developed on the well surfaces of 12-well plates and on hydroxyapatite (HA) coupons, and then exposed to IP6. In addition, selected HA coupons were preconditioned with IP6 before biofilm development. IP6 demonstrated bactericidal effects and altered the metabolic activity of biofilm cells. Confocal laser-scanning microscopy showed that IP6 caused significant and rapid reduction in live biofilm cells. At sublethal concentrations, IP6 did not alter the expression of tested virulence genes except for C. albicanshwp1, the expression of which was upregulated but not reflected by a change in hyphal transformation. IP6-preconditioned HA coupons led to extensive inhibition of dual-species biofilm formation. The results of this study highlight for the first time the antibiofilm inhibitory properties of IP6 and the potential for its exploitation in several clinical applications.Root canal infections are associated with biofilms and are treated with chemical irrigants with a high success rate. However, treatment failure does arise, which is attributed primarily to resistance exhibited by biofilms. Currently used irrigants in root canal treatment have disadvantages, and there is therefore a need for more biocompatible alternatives with antibiofilm properties to reduce root canal treatment failure and complications. The aim of this study was to evaluate the IMPORTANCE Root canal infections are biofilm associated, and despite mechanical and chemical treatment procedures, infection recurrence occurs, and this is likely due to the high tolerance of associated biofilms to antimicrobials. The currently used treatment agents have several disadvantages, which necessitates the search for new improved agents. In this study, the natural chemical phytic acid was found to exhibit antibiofilm activity against established mono and dual mature biofilms over a short contact time. Most importantly, phytic acid was found to cause significant inhibition of dual-species biofilm formation when used as a surface preconditioning agent. The findings of this study identified a novel use of phytic acid as a potential antibiofilm agent that can be used in several clinical applications. Enterococcus faecalis and Candida albicans are frequently recovered from persistent infections where endodontic treatment has failed  is a natural agent that has been proposed as a potential root canal irrigant because of its ability to remove the smear layer and its higher biocompatibility than EDTA . In genesruption , 31. Zhohimurium . In a rel growth . Howeverl growth . However570]; biomass) were calculated. AlamarBlue is a redox indicator, and metabolically active cells reduce the blue color resazurin to resorufin, which has a fluorescent pink color. The inability of cells to reduce alamarBlue is indicative of reduced metabolic activity. Generally, a significant reduction in mean RFU in treated groups compared to untreated groups occurred  significantly reduced the biomass of single-species E. faecalis ATCC 29212 (P = 0.017) biofilms and dual-species biofilms (P = 0.024). For E. faecalis  biofilms, significant biomass reduction was seen at lower IP6 concentrations   assays, respectively. The means of relative fluorescence units  and absorbance  . After cregrowth . Since rregrowth  and b. FCC 29212 .E. faecalis ATCC 29212 mono-species biofilms, IP6 at 1.25%, 2.5%, and 5% led to a significant reduction in log10 (CFU) values compared with untreated controls  of 96.79%, 98.98%, and 99.70%, respectively . For dual-species biofilms, 1.25%, 2.5%, and 5% IP6 led to significant reductions in E. faecalis log10 (CFU) values . The presence of both E. faecalis and C. albicans in biofilms did not affect IP6 efficacy against E. faecalis. However, in all cases of IP6 treatment , no reduction in C. albicans ATCC 90028 log10 (CFU) values occurred . For controls , with re) values , with reoccurred . Live bioccurred . The biooccurred . The samoccurred . The reloccurred . Comparioccurred . CLSM imE. faecalis and C. albicans was investigated by CLSM and scanning electron microscopy (SEM). CLSM quantitative image analysis revealed significant reductions in total and live-cell biomass of biofilms formed on HA coupons preconditioned with 2.5% IP6   . This wactively)  and b. HE. faecalis ATCC 29212 and C. albicans ATCC 90028 in biofilms following treatment with 0.16% IP6 was quantified. All analyzed E. faecalis genes demonstrated no significant changes in expression levels for IP6-treated biofilms compared to untreated biofilms. The expression of C. albicans genes was unchanged in treated biofilms with the exception of hwp1 gene expression, which was upregulated   .IP6 is a natural compound with high biocompatibility that has been proposed as an alternative chelating agent to EDTA for root canal smear layer removal . Severalin vitro biofilms of E. faecalis and C. albicans were tested, as these species are frequently associated with persistent endodontic infection  and incubated aerobically at 37°C for 24 h. Candida albicans was cultured on Sabouraud dextrose agar (SDA) at 5% CO2 and 37°C for 48 h.faecalis , and CanE. faecalis ATCC 29212 was mixed with an equal volume of a 1.0 McFarland standard of C. albicans ATTC 90028. One milliliter of microbial suspension was added to each well of the 12-well plate. The biofilm of the oral strain of E. faecalis was grown statically for 48 h, while biofilms of E. faecalis ATCC 29212 and dual biofilms were grown for 72 h. Biofilms were cultured aerobically at 37°C, and the medium was changed every 24 h.Mono- and dual-species biofilms were developed on the well surfaces of 12-well plates that had been preconditioned with Dulbecco’s Modified Eagle’s Medium (DMEM) for 24 h. Briefly, microbial colonies were used to prepare suspensions in tryptone soya broth (TSB), which were then incubated at 37°C with agitation (125 rpm) in a shaker incubator for 3 to 4 h. For mono-species biofilms, bacterial suspensions were adjusted to a 0.5 McFarland standard using a DensiCHEK meter (bioMérieux). For the dual-species inoculum, a 1.0 McFarland standard of After biofilm formation, spent medium was discarded, and biofilms were washed twice with phosphate-buffered saline (PBS). One milliliter of TSB and 1 mL of IP6 (Sigma), with a concentration range of 40% to 0.31% in water, were added to designated wells. Untreated biofilm and blank controls received 1 mL of TSB and 1 mL of sterile distilled water. The biofilms were incubated aerobically for 24 h at 37°C. After IP6 treatment, test solutions were removed, and the biofilms were washed twice with PBS. Two mL of 10% alamarBlue solution was added to each well, and the plates were incubated for 1 h at 37°C. Fluorescence of the wells was then measured using a microplate reader . To assess total biomass, alamarBlue suspension was discarded, and a 2-mL volume of 0.1% crystal violet (CV) was added and incubated for 10 min at room temperature. The CV was removed, and the wells were washed twice with distilled water. Excess water was removed, and the plates were allowed to air dry for 30 min. Two milliliters of 70% ethanol was added to each well to elute bound CV, and the plate was incubated at room temperature for 10 min. The absorbance of the wells was then read using a microplate reader at 570 nm.After treatment of the biofilms with IP6 for 24 h, test solutions were removed from the wells, which were then washed twice with PBS. Two milliliters of sterile TSB was then added to all wells, followed by 18 to 24 h of aerobic incubation at 37°C. After incubation, regrowth was assessed by reading the absorbance of the wells with a microplate reader  at 600 nm.The IP6 concentrations that allowed bacterial regrowth were used to study the effects of IP6 on bacterial cell count reduction. After treatment of the biofilms with IP6 for 24 h, the solution was removed from the wells, which were then washed twice with PBS. The biofilms were then detached by scraping and vigorous pipetting, and subsequent enumeration of viable cells was performed.E. faecalis ATCC 29212 bacterial suspension. One milliliter of a 0.5 MacFarland standard E. faecalis ATCC 29212 and 1 mL of a 0.5 MacFarland standard C. albicans ATTC 90028 were inoculated for dual-species biofilms. The CDC Biofilm Reactor was incubated as a batch system for 18 to 24 h at 37°C while placed over a stirrer plate for baffle stirring to produce shear stress. After incubation, the biofilm reactor were connected with a carboy containing sterile 1 g/L TSB to run a continuous flow system for an additional 48 h at a flow rate of 1.1 mL/min. The biofilms formed on HA coupons were then treated with three concentrations of IP6  for 5 min, and the antibiofilm activity of IP6 was assessed as outlined below.Center for Disease Control (CDC) Biofilm Reactors were assembled, and HA coupons were positioned in designated sites in the support rods before sterilization. The system flask was aseptically filled with 325 mL of 3 g/L TSB. For mono-species biofilms, the CDC Biofilm Reactor was inoculated with 1 mL of a 0.5 MacFarland standard A single tube method  was usedwww.comstat.dk) to quantify live and dead biomass. CLSM micrographs were generated using Imaris viewer software (9.9.1 version).After biofilm development in the CDC Biofilm Reactor, the rods were washed and treated as described above. Biofilms were stained with live/dead BacLight bacterial viability mixture  for 10 min at room temperature and immediately analyzed by CLSM. Five random fields of view were imaged for each coupon using an LSM880 Airyscan microscope . Quantitative image analysis was performed using Comstat 2.1 . For CLSM, coupons were processed as described above, and for SEM, coupons were washed with PBS and placed immediately in 12-well plates containing 2.5% glutaraldehyde and incubated overnight. After incubation, the biofilms were dehydrated in 2 mL of an ethanol solution series  for 10 min at each concentration. Dehydration with 100% ethanol was repeated twice. The samples were then sputter coated with gold and imaged using a Tescan VAGA SEM system at 5 to 10 kV.To assess the effect of preconditioning HA coupons with IP6 on biofilm formation, coupons were soaked in either 2.5% IP6 or sterile distilled water for 18 h under aseptic conditions. The rods were removed and then placed in an assembled sterile CDC Biofilm Reactor containing TSB. The CDC Biofilm Reactor was inoculated with 1 mL of a 0.5 McFarland standard level of both −ΔΔCt method. Target genes and primer sequences are provided in Tables S1 and S2 in the supplemental material.The effect of 0.16% IP6 on gene expression of 72-h-old dual-species biofilms developed in 12-well plates, as described above, was assessed. Developed biofilms were challenged with 0.16% IP6 for 24 h. The test solution was removed, and the wells were washed twice with PBS. One milliliter of RNAprotect bacteria reagent (Qiagen) was added and left at room temperature for at least 10 min. The biofilms were detached by scraping and vigorous pipetting. The resulting suspension was processed for RNA extraction using a Qiagen kit, and RNA was then reverse transcribed as per the manufacturer’s protocol (Qiagen). Quantitative PCR was performed using SYBR green mix (Bio-Rad) under the following thermal cycling conditions: 95°C for 2 min, 40 cycles of 95°C for 5 s, and 60°C for 20 s. Gene expression was normalized to the expression of housekeeping genes and the gene expression of treated biofilms was calculated relative to untreated biofilms (control) using the 2Candida fungal units (yeast and hyphae) was determined using ImageJ software version 1.48.To study the effect of IP6 on hyphal production, 72-h dual-species biofilms were prepared and treated with 0.16% IP6 as described above. After treatment, the medium was retrieved, and the biofilm was washed twice with PBS and fixed with 4% paraformaldehyde at room temperature for 1 h. The wells were washed with PBS and stained with 0.2% calcofluor white for 30 min at room temperature. Images were obtained using an Olympus IX53 fluorescence inverted microscope. The proportion of hyphae relative to total t test was used to compare differences between any two groups. A Kruskal-Wallis test or ordinary one-way analysis of variance (ANOVA) was used to compare more than two groups. A P value of less than 0.05 was considered significant. Relative fold change in gene expression analysis was performed using a one-sample t test compared to a hypothetical mean of 1.Statistical analysis was performed using GraphPad Prism 9.4.0. A parametric