Tracheobronchial foreign body is uncommon in adults. Among foreign body aspirations, tooth and dental prosthesis aspiration is a very rare condition. In the literature, dental aspiration is generally found as a case report and there is no single-center case series. In this study, we aimed to present our clinical experience in 15 cases with tooth and dental prosthesis aspiration.Data from 693 patients who presented to our hospital for foreign body aspiration between the years 2006 and 2022 were analyzed retrospectively. Fifteen cases who aspirated tooth and dental prostheses as foreign bodies were included in our study.Foreign bodies were removed by rigid bronchoscopy in 12 (80%) cases and fiberoptic bronchoscopy in 2 (13.3%) cases. In one of our cases, foreign body was expected with cough.When evaluated in terms of foreign body, partial upper anterior tooth prosthesis in 5 (33.3%) cases, partial anterior lower tooth prosthesis in 2 (13.3%) cases, dental implant screw in 2 (13.3%) cases, lower molar crown in 1 (6.6%) case, lower jaw bridge prosthesis in 1(6.6%) case, upper jaw bridge prosthesis in 1(6.6%) case, broken tooth fragment in 1(6.6%) case, upper molar tooth crown coating in 1(6.6%) case and upper lateral incisor tooth in 1(6.6%) case were observed.Dental aspirations can also occur in healthy adults. Anamnesis is the most important factor in diagnosis and diagnostic bronchoscopic procedures should be performed in cases where adequate anamnesis cannot be obtained. Tracheobronchial foreign body aspiration is one of the life-threatening emergencies. It is most common in children aged 1–3 years. Tracheobronchial foreign body aspirations are very rare in adults. However, it has been reported that there is a significant increase in the frequency of foreign body aspiration in adults after the age of fifty years . In addiData from 693 patients who presented to our hospital for foreign body aspiration between the years 2006 and 2022 were analyzed retrospectively. Fifteen cases who aspirated tooth and dental prostheses as foreign bodies were included in our study. The cases were evaluated in terms of age, gender, physical examination, medical history, duration of dental prosthesis, symptoms, admission time to the hospital, radiological examination and radiological findings, nature and localization of foreign body, type of anesthesia, bronchoscopic procedure type and complications. Chest X-rays and thorax computerized tomography (CT-scans) were used in the radiological evaluation of the cases. Hemogram, coagulation factors and routine biochemistry tests were performed in all cases.A rigid and flexible bronchoscope was used to remove foreign bodies. Oral feeding was stopped 6–8 h before all bronchoscopic procedures. All bronchoscopic procedures were performed under operating room conditions. Aerosol lidocaine (max 8.2 mg/kg) as a local anesthetic and intravenous midazolam (0.06–0.07 m/kg) for sedation were administered before a flexible bronchoscopy procedure . Rigid bronchoscopy  procedures were performed under general anesthesia. In rigid bronchoscopy, foreign bodies were removed using crocodile and peanuts forceps. Foreign bodies were removed using basket forceps in flexible bronchoscopy.p-value < 0.05 was considered as statistically significant.Statistical analysis was done with IBM SPSS 20.0  package program. Normal distribution was evaluated with the Shapiro–Wilk test. Numerical variables were given as mean ± standard deviation. Categorical variables were given as frequency (percentage). Comparison of continuous variables between groups was carried out using Mann–Whitney U test. All statistical analyses were carried out with 5% significance and a two-sided p = 0.06).A total number of 15 cases were included in our study. In terms of gender distribution, 5 (33.3%) of our cases were female and 10  were male. The mean age of the cases was 58.10 ± 9.07 (range of 32 to 71) years. The mean age was 63.43 ± 10.13 (range of 47 to 82) years in males and 47.00 ± 10.36 (range of 32 to 59) years in females. No statistically significant difference was found between the male and female groups in terms of age distribution  had hypertension, 6 (40%) had coronary artery disease, 5 (33.3%) had diabetes mellitus, 4 (26.6%) had obstructive sleep apnea syndrome, 4 (26.6%) had chronic obstructive pulmonary disease, 4 (26.6%) had bruxism, 3 (20%) had asthma, 3 (20%) had depression, 3 (20%) had alcoholism and 1 (6.6%) had myasthenia graves  cases, decrease in breathing sounds in 9 (60%) cases, stridor in 6 (40%) cases, decreased saturation in 4 26.6%) cases and there was no finding in 4 (26.6%) cases  hours. The diagnosis of foreign body was made three weeks later in one patient who was in the intensive care unit and aspirated his tooth after trauma. A case who aspirated the molar tooth crown did not remember the time of aspiration.A chest X-ray was performed for radiological examination in all cases. Computerized tomography of thorax was performed in 9 (60%) cases. On radiological evaluation, all of the foreign bodies were detected directly.Foreign bodies were removed by rigid bronchoscopy in 12 (80%) cases and fiberoptic bronchoscopy in 2 13.3%) cases  cases Fig. , partial3.3% caseForeign bodies were evaluated in terms of location as follows: 9 foreign bodies (60%) were in the right main bronchus, 4 (26.6%) in the left main bronchus and 2 (13.3) in the right lower lobe bronchus.As a complication, mucosal injury in the trachea during the removal of the foreign body in 3 (20%) cases and hoarseness due to edema in the vocal cords in 2 (13.3%) cases occurred. In the controls performed one week after bronchoscopy, no finding was found in the physical examination of the cases.In our study, the rate of tooth and dental prosthesis aspiration was found to be 2.2%.Tracheobronchial foreign body aspirations are less common than gastrointestinal foreign bodies. However, foreign bodies in the tracheobronchial system cause higher mortality and morbidity than the gastrointestinal system. While the mortality rate in tracheobronchial system foreign bodies is approximately 5–7%, this rate is around 0.85% in gastrointestinal tract foreign bodies , 5. AspiForeign body aspirations are evaluated in terms of gender, it is more common in men . The majDental caries occurs in elderly patients due to deterioration in oral hygiene. Although a decrease in the prevalence of tooth decay has been observed worldwide, untreated tooth decay in permanent tooth is quite common and occurs in approximately 35% of the world's population . Tooth dMaxillofacial trauma and emergency intubations are among the predisposing factors in dental aspirations , 12. ForDuring dental treatments, some dental tools or screws can be aspirated. In our two cases, the dental screw was aspirated. Therefore, some preventive measures have been recommended to dentists. These include the use of an intraoral protective cover, adequate secretion cleaning so that patients do not need to breathe deeply, and tying some prosthesis parts with a thread . HoweverDetailed anamnesis is very important in the diagnosis of all foreign body aspirations. Sudden onset of cough and respiratory distress in the patient's history is diagnostic for foreign body aspiration. The diagnosis of foreign body aspiration may be delayed in cases where adequate anamnesis cannot be obtained. Delayed diagnosis in foreign body aspirations may cause recurrent lung infections, lung abscesses, bronchiectasis, hemoptysis, and formation of granulation tissues that can be confused with lung cancer . BronchiBronchoscopic methods are most commonly used in the treatment of foreign body aspirations. Although the most commonly used method is rigid bronchoscopy, fiberoptic bronchoscopy can be used in the treatment of foreign body aspiration. The advantages of fiberoptic bronchoscopy are that it can be performed under local anesthesia and sedation. Disadvantages of fiberoptic bronchoscopy are that the forceps are quite small, there is a risk of foreign body escaping back into the airway, and small foreign bodies can be removed . In our In conclusion, tracheobronchial aspiration of tooth and dental prostheses is quite rare. Although foreign body aspirations are frequently seen in adults with advanced age, dementia, alcoholism, neurological diseases and maxillofacial trauma, it should be kept in mind that dental aspirations can also occur in healthy adults. Anamnesis is the most important factor in diagnosis, and diagnostic bronchoscopic procedures should be performed in cases where adequate anamnesis cannot be obtained.


Psoriasis is a chronic autoinflammatory skin disease that affects 2% of the population worldwide.Ocular adverse events associated with clobetasol, methotrexate, and adalimumab that were reported to the Federal Drug Administration (FDA) Adverse Event Reporting System (FAERS) Database were gathered and assessed.In total, 3466 total adverse events associated with clobetasol were reported to FAERS. Of these events, 19 (0.55%) were cataracts and 22 (0.63%) were glaucoma. A total of 7359 adverse events reported were associated with methotrexate for patients being treated for dermatologic conditions only, of which 140 (1.91%) were cataracts and 382 (5.19%) were glaucoma. A total of 41,737 adverse events reported were associated with adalimumab for patients being treated for dermatologic conditions only, of which 198 (0.47%) were cataracts and 74 (0.18%) were glaucoma.P > 0.05) or adalimumab . Patients being treated for psoriasis with clobetasol had a lower rate in the incidence of glaucoma when compared to those being treated with methotrexate  and no statistically significant difference when compared to adalimumab . A recent review of all case reports regarding TCS associated ocular adverse events by Daniel et al.According to the FAERS data, patients treated with clobetasol have an overall low incidence of cataracts (0.55%) and glaucoma (0.63%) as adverse events. Worldwide, the incidence of cataracts is approximately 2.5% in those aged of 40–49 years, 6.8% in those aged of 50–59 years, 20.0% in those aged of 60–69 years, 42.8% in those aged of 70 to 79 years, and 68.3% in those aged greater than 80 years.Of note, it is essential to educate patients about avoiding application of clobetasol to the face, as it can cause perioral dermatitis/steroid rosacea.Dr. Wang and Dr. Jorizzo have no conflicts of interest relevant to the content of the submission. This work has not been previously published.


Although perfusion magnetic resonance imaging (MRI) is widely used to identify pseudoprogression, this advanced technique lacks clinical reliability. Our aim was to develop a parameter assessing the hypervascularized fraction of glioblastomas based on volume analysis of dynamic susceptibility contrast-enhanced MRI and evaluate its performance in the diagnosis of pseudoprogression.Patients with primary glioblastoma showing lesion progression on the first follow-up MRI after chemoradiotherapy were enrolled retrospectively. On both initial and first follow-up MRIs, the leakage-corrected cerebral blood volume (CBV) maps were post-processed using the conventional hot-spot method and a volume method, after manual segmentation of the contrast-enhanced delineated lesion. The maximum CBV (rCBVmax) was calculated with both methods. Secondly, the threshold of 2 was applied to the CBV values contained in the entire segmented volume, defining our new parameter: %rCBV>2. The probability of pseudoprogression based on rCBVmax and %rCBV>2 was calculated in logistic regression models and diagnostic performance assessed by receiving operator characteristic curves.Out of 25 patients, 11 (44%) were classified with pseudoprogression and 14 (56%) with true progression based on the Response Assessement in Neuro-Oncology criteria. rCBVmax was lower for pseudoprogression  on early follow-up MRI. %rCBV>2, was lower for pseudoprogression on both initial  and early follow-up MRIs . On early follow-up MRI, %rCBV>2 had the largest area under the curve for the diagnosis of pseudoprogression: 0.909 [0.725–0.986].The fraction of hypervascularization of glioblastomas as assessed by %rCBV>2 was lower in tumours that subsequently developed pseudoprogression both on the initial and early follow-up MRIs. This fractional parameter may help identify pseudoprogression with greater accuracy than rCBVmax. Sensitivity and specificity were calculated for cut-off values determined with the Youden index.For inter-observer reproducibility, the intraclass correlation coefficient (ICC) of quantitative measurements of %rCBV>2 and a kappa score for pseudoprogression diagnosis were calculated. The ICC was calculated with a two-way random model, with single measure consistency and reported with their 95% confidence interval. Agreement was considered using standard guidelines .All tests were bilateral and a p-value of 0.05 indicated statistical significance. Statistical analysis was performed with a biostatistician (A.G.) using commercially available software SAS Version 9.4  and STATA version 14.2 .Among the 25 patients included, 11 were classified as having pseudoprogression (44%) and 14 as having tumour progression (56%). The entire cohort had a median age of 62 years at the time of diagnosis [range 42–82]. There were 11 women (44%) and 14 men (56%) and median survival was 17.1 months [5.4–47.5]. The patients in the pseudoprogression group were significantly younger  and had a longer survival  than patients in the true progression group. The MGMT promoter methylation status was positive in 55% of patients with pseudoprogression and in 36% of patients with tumour progression (p = 0.435). The basic patient characteristics are summarised in The median rCBVmax values of untreated glioblastomas calculated with the two different methods were comparable: 9.8 [8.9–10.8] with the hot-spot and 10.2 [9.6–10.7] with the volume method. The two post-processing methods had a good level of concordance: the mean difference between the calculated values was -0.48, with the highest variations for extreme values and the Spearman correlation coefficient differed significantly from 0  .On the initial MRI, with the hot-spot method, the median rCBVmax was not different between patients with subsequent pseudoprogression and tumour progression, 9.1 [7.1–12.4] vs. 10.1 [9.6–13.4] (p = 0.14), respectively. A similar finding was obtained with the volume method, namely 9.8 [8.7–10.6] vs. 10.5 [9.5–11.1] (p = 0.27), respectively . The fraOn CRT+1 MRI, with the hot-spot method, rCBVmax was lower for pseudoprogression than for tumour progression, 3.4 [2.4–7] vs. 7.6 [4.9–9.6], respectively (p = 0.03) but with substantially overlapping values . CalculaThe ROC curve comparison showed a tendency for better diagnostic accuracy for %rCBV>2 (AUC = 0.909) compared to rCBVmax calculated with the hot-spot  or the volume method . When %rCBV>2 was under 41.5%, pseudoprogression could be diagnosed with 100% sensitivity and 71% specificity . Fig 5 iThe inter-observer agreement for quantitative measurement of %rCBV>2 was excellent (ICC = 0.88 [0.75–0.95]) as for pseudoprogression diagnosis with the 41.5% threshold .Including the central necrotic non-enhancing component of the lesions could have led to an underestimation of %rCBV>2 and a misdiagnosis of pseudoprogression, especially in case of highly necrotic lesions. On CRT+1 MRIs, we secondly segmented those necrotic parts and excluded them from the volume analysis. Pseudoprogression mean %rCBV>2 was 23.3 ± 13.7 and tumour progression mean %rCBV>2 was 61.1 ± 16.7. The new AUC for %rCBV>2 was 0.883 [0.692 to 0.976] (p = 0.507), showing no significant advantage in excluding necrosis from the analysis. Two of the 3 patients with tumour progression who were misdiagnosed as having pseudoprogression still displayed a %rCBV>2 under the 41.5% threshold .From the volume analysis of DSC-PWI, we developed a parameter that assessed the fraction of hypervascularized tissue of an entire contrast-enhanced cerebral tumour: %rCBV>2. This fractional parameter helped to differentiate subsequent pseudoprogression from true progression of treated glioblastomas, even on the initial MRI  and with a higher confidence level on the early follow-up MRI performed 1 month after CRT . On the early follow-up MRI, when a lesion had a %rCBV>2 under 41.5%, pseudoprogression could then be diagnosed with 100% sensitivity and 71% specificity.Evaluation of the fraction of an entire contrast-enhanced lesion displaying a rCBV greater than 2, allowed us to assess the proportion of hypervascularization within this lesion. We assumed that this parameter would provide a better indication of the hypervascularization of heterogeneous tumours such as glioblastomas than measurements of a few selected points using the conventional hot-spot method. On the initial MRI, the conventional parameter rCBVmax was no different for glioblastomas that would subsequently develop pseudoprogression or tumour progression after CRT. However, the fractional parameter, %rCBV>2, was lower in the pseudoprogression group, potentially indicating a difference in the intrinsic neoangiogenesis and aggressiveness of the naive tumour. %rCBV>2, unlike rCBVmax, may be able to depict tumour sub-groups in an apparently homogeneous cohort. Those sub-groups could be linked to the genetic status of tumours, such as the MGMT status or other mutations, associated with a better therapeutic response and prognosis , 22. In As with previous studies, on early follow-up MRI (CRT+1), calculation of rCBVmax with the hot-spot method enabled us to differentiate between pseudoprogression and tumour progression groups , 23. HowA similar parameter, the perfusion MRI-fractional tumour burden (pMRI-FTB), defined as the percentage of tumour voxels relative to total lesion mask voxels, has already been correlated with the overall survival of progressive glioblastomas and used to differentiate treatment effect from tumour recurrence , 25. Theet al. showed that freeform ROIs that encompassed the whole tumour had a good diagnostic accuracy in differentiating low grade from high grade tumour [Unlike for pMRI-FTB and some previous studies , 24, we e tumour . In accoThis study has some limitations. First, the retrospective and limited number of patients, which is common in most studies on pseudoprogression. To ensure a homogenous cohort, we used restrictive inclusion and exclusion criteria. For example, all perfusion data used were acquired on 3T systems since rCBV can be over-evaluated at 3T in comparison to 1.5T . HoweverThe fraction of hypervascularization in glioblastomas assessed by our fractional parameter %rCBV>2 was lower in patients that will subsequently develop pseudoprogression compared to true progression, on both initial and early follow-up MRIs. This parameter seems to better represent the heterogeneity of these enhancing lesions and better identify pseudoprogression than rCBVmax, conventionally calculated with the hot-spot method. In our cohort pseudoprogression could have been diagnosed on early follow-up MRI with 100% sensitivity and 71% specificity when %rCBV>2 was under 41.5%.


Glycosylated haemoglobin (HbA1c) is considered reliable for diagnosing and monitoring diabetes mellitus (DM). It also indicates cardiovascular complications related to DM. However, only a few studies have been conducted on this topic.We conducted a cross-sectional study to investigate the association between newly diagnosed hypertension and HbA1c among non-diabetic Sudanese adults. The sociodemographic characteristics of the participants in the sample were gathered using a questionnaire, and HbA1c was measured using an Ichroma machine.2. One hundred and fifteen (29.9%) participants presented newly diagnosed hypertension. The results of the multivariate analysis showed that age ; BMI ; HbA1c levels  was positively associated with newly diagnosed hypertension. For an HbA1c level of 5.0% or more, the sensitivity and specificity of newly diagnosed hypertension were 91.3% and 28.2%, respectively . Participants who presented HbA1c levels of 5.0% or more were found to be at higher risk for newly diagnosed hypertension .Three hundred and eighty-four healthy participants were enrolled in this study. The median (interquartile range [IQR]) age was 56.0 (14.0) years, and 72.1% of the participants were female. The median (IQR) body mass index (BMI) was 31.2 (8.7) kg/mThe results of this study indicated a high prevalence of newly diagnosed hypertension, and HbA1c levels were positively associated with newly diagnosed hypertension. The incidence of non-communicable diseases has increased globally, especially in African countries . Non-comIn 2019, the global prevalence of diabetes mellitus (DM) was estimated to be 9.3% (463 million people) and was expected to rise to 10.2% 578 million) and 10.9% (700 million) by 2030 and 2045, respectively ) where applicable. A univariate analysis was performed, in which newly diagnosed hypertension was the dependent variable. The independent variables were age, gender, employment, education, BMI and HbA1c. Multicollinearity (variance inflation factor pf less than four) was checked but not detected. A logistic regression analysis was applied to the variables if their univariate 2 , BMI  and HbA1c levels  were found to be positively associated with newly diagnosed hypertension. There was no significant association between marital status, level of education and newly diagnosed hypertension (Table P ˂ 0.001) .For HbA1c levels of 5.0% and above, the sensitivity and specificity of the diagnosis of newly diagnosed hypertension were 91.3% and 28.2%, respectively . Using a cut-off point for HbA1c levels of 5.0% or more, the sensitivity and specificity of the diagnosis of hypertension were calculated as 91.3% and 28.2%, respectively. These results are in line with those of a prospective cohort study that includes 9,603 middle-aged American participants, where elevated HbA1c levels (even without a prior diagnosis of DM) were associated with hypertension among diabetic as well as non-diabetic individuals . FurtherIn contrast to our findings, a recently published study demonstrated that HbA1c in diabetic patients was not correlated with arterial stiffness; however, the concomitant hypertension was a significant potential risk for arterial stiffness . FurtherInsulin resistance is the main pathophysiological factor behind the increased risk of developing both T2DM and hypertension . In addiHbA1C is a significant predictor of newly diagnosed hypertension among healthy adult non-diabetic Sudanese. Hence, it can be used to modify or prevent hypertension among those who have considerably high HbA1c levels.Our study has the following limitations. The causes and the effects of the results could not be inferred, as this was a cross-sectional study. Only a single measurement of HbA1c at baseline was available, which may have resulted in misclassification. Other causes of secondary hypertension cannot be excluded. Moreover, other factors, such as dyslipidemia, were not assessed. Also the high number of females participant which may be due to the availability of females at home during the data collection.


The current study evaluated the effect of pomegranate peel-based edible coating on chicken nuggets in order to develop a functional and safe product, high in nutritional value. For this purpose, 2,2-diphenyl-1-picrylhydrazyl (DPPH) and total phenolic content (TPC) assays were performed to check the potential antioxidant activity of chicken nuggets; microbial control, including total aerobic count and coliforms population, was performed for quality and safety purposes; and thiobarbituric acid reactive substances (TBARS) and peroxide value (POV) were performed to determine the oxidative stability of chicken nuggets. Different treatments were applied at different storage periods . The higher value of total aerobic count (5.09 ± 0.05 log CFU/g) and coliforms (3.91 ± 0.06 log CFU/g) were obtained for the uncoated samples, while the lower population was enumerated in the combination of sodium alginate (SA) and pomegranate peel powder (PPP). However, DPPH (64.65 ± 2.15%) and TPC (135.66 ± 3.07 GAE/100 g) values were higher in the coated chicken nuggets (SA (1.5%) and PPP (1.5%)) and lowest in the control samples. The higher value of TBARS (1.62 ± 0.03 MDA/kg) and POV (0.92 ± 0.03 meq peroxide/kg) were observed in the uncoated chicken nuggets. In the Hunter color system, L*, a*, and b* peak values were determined in the coated chicken nuggets with SA (1.5%) + PPP (1.5%) at the 21st day of storage. The uncoated chicken nuggets had different sensory characteristics  compared to the coated samples. Conclusively, coating based on the combination of SA (1.5%) and PPP (1.5%) increased the quality, safety, and nutritional properties of chicken nuggets. Escherichia coli O157:H7 and Listeria monocytogenes) has also spurred greater interest in finding innovative technologies to control infectious growth in foods. These technologies maintain quality, safety, and freshness [Microbial contamination affects the safety and quality characteristics of foods, and may occur at any stage, during production, transport, processing, meal preparation, domestic storage, and retail trade ,2. Food-reshness . Antimicreshness . SpoilagAntimicrobial edible coating is among the novel technologies to control the bacterial development and improves protection, while delays degeneration of essence, fish and pullet products . EssentiPunica granatum) belongs to the Punicaceae family and is commonly called ‘’ponus’’ and ‘’granatus’’ that is consequent from the Latin words. The wrapping of the pomegranate characterizes practically 26–30% of the fruit. The polyphenols including flavonoids , hydrolysable tannins , and total antioxidant capacity are high in this portion of pomegranate. These chemical compounds are present in the pomegranate peel and juice, and justify the 92% of the antioxidant activity related with the fruit [Pomegranate  separately or in combination. At the 0th, 7th, 14th, and 21st days of storage intervals, DPPH, TPC, TBARS, POV, Hunter color values , and sensory analyses were carried out to investigate the functionality, quality, and oxidative stability of chicken nuggets.The total aerobic count in uncoated and coated samples of chicken nuggets during storage are given in p < 0.05) by the coating formulations and storage time.The microbial population for total aerobic count ranged between 3.05 ± 0.01 to 5.09 ± 0.05 log CFU/g. On the 21st day of storage, the microbial population for total aerobic count ranged from 3.69 ± 0.06 to 5.09 ± 0.05 log CFU/g. During storage at refrigerated temperature, a slight changes in total aerobic count was determined in the coated samples. An extreme value of total aerobic count (5.09 ± 0.05 log CFU/g) was recorded in uncoated sample at 21st day of storage, whereas the least value of total aerobic count (3.05 ± 0.01 log CFU/g) was obtained with the coating of SA (1.5%) and PPP (1.5%) on 0th day. The present findings showed that the total aerobic count was affected significantly  in the uncoated as compared to coated chicken nuggets.The microbial population of coliforms ranged from 1.46 ± 0.04 to 2.42 ± 0.06 log CFU/g in the beginning of the experiment. On the 21st day of storage, the microbial population for coliforms ranged from 1.74 ± 0.02 to 3.91 ± 0.06 log CFU/g. Minor reinstate in coliforms was observed in the coated samples  during refrigerated storage. A high value of coliforms (3.91 ± 0.06 log CFU/g) was recorded on the 21st day of storage, although a minor coliforms population of 1.46 ± 0.04 log CFU/g was obtained for the coating of SA (1.5%) and PPP (1.5%) on 0th day. Therefore, present results showed that the coliforms population increased significantly (p > 0.05) difference between SA and PPP during refrigerated storage. However, the amount of PPP (1.5%) and SA (1.5%) + PPP (1.5%) showed a good efficiency in preventing lipid oxidation in the coated chicken nuggets.Thiobarbituric Acid Reactive Substances control is another methodology that is normally used as a guide of rancidity of fatty foods during storage . The TBAPOV basically measures the number of peroxides that develop in meat products due to auto-oxidation. During processing, the oxidation process starts due to the auto-oxidation of unsaturated fats. The results of auto-oxidation produce off essences and off-odors that directly affect the quality of products.p < 0.05) in the uncoated chicken nuggets samples with respect to storage time.The peroxide values of uncoated, SA (1.5%), PPP (1.5%), and SA (1.5%) + PPP (1.5%) formulations at refrigerated temperature (4 ± 1 °C) are shown in p < 0.05) on the coated (SA (1.5%) + PPP (1.5%)) chicken nuggets compared to the uncoated ones, during storage under refrigeration. This finding is due to the high amount of phenolic compounds in the coated samples, given that the coating materials used contain rich phenolic compounds. For instance, sodium alginate contains glucuronic acid and monotonic acid that develop a layer. This layer aids in the reduction of microorganisms population and prevents hydration of the product.The TPC of uncoated, SA (1.5%), PPP (1.5%), and SA (1.5%) + PPP (1.5%) at refrigerated temperature are given in p < 0.05) affected by the coating formulations at different storage time.The antioxidant activity of chicken nuggets based on the DPPH assay in regard to the treatments , PPP (1.5%), and SA (1.5%) + PPP (1.5%) formulations) and storage time is shown in The Hunter color scale is a quality parameter of food products on the basis of their developed color. During coating, the color of products depends upon the ingredient that are used for coating. The color values of uncoated, SA (1.5%), PPP (1.5%), and SA (1.5%) + PPP (1.5%) formulations at refrigerated temperature are shown in p < 0.05) affected by non-coating and storage time. The L* values of chicken nuggets increased with respect to the coating formulations and storage time. The coated samples had higher L* values as compared to the uncoated samples. Increment in the L* values may be due to the high antioxidants content in the used coating materials. Higher a* values (14.87 ± 0.62) were obtained for the chicken nuggets coated with SA (1.5%) + PPP (1.5%) at 21st day of storage, while the minimum a* values (11.36 ± 0.45) were obtained for the uncoated chicken nuggets on 0th day. However, the present results showed that a* values were affected significantly (p < 0.05) by non-coating and storage time. The a* values of chicken nuggets were significantly different. The coated samples had higher a* values as compared to the uncoated samples. These differences are probably due to the coating material used (SA and PPP). Higher b* values (11.25 ± 0.34) were obtained for the chicken nuggets coated with SA (1.5%) + PPP (1.5%) on the 21st day of storage, while the minimum b* values (9.15 ± 0.34) were obtained for the uncoated chicken nuggets on 0th day. During storage, the b* values of coated chicken nuggets were found to be enhanced as compared to the uncoated samples. The increments in the present results may be due to the oxidation process that starts during storage. Moreover, results showed that b* values of coated chicken nuggets increased significantly (p < 0.05) with respect to the non-coating samples, due to the different composition of the used coating materials.Higher L* values (62.36 ± 2.15) were obtained for the chicken nuggets coated with SA (1.5%) + PPP (1.5%) at 21st day of storage, whereas the lower L* values (58.12 ± 1.06) were obtained for the uncoated chicken nuggets on 0th day. Present findings showed that L* values were significantly (p < 0.05) with respect to the coating formulations and storage time.The pH values of uncoated, SA (1.5%), PPP (1.5%), and SA (1.5%) + PPP (1.5%) formulations during refrigerated storage are shown in p < 0.05) with respect to the coated chicken nuggets and storage time.The appearance score values of chicken nuggets for the different treatments are shown in p < 0.05) with respect to the uncoated chicken nuggets at different storage time.The mean ± SD texture score values of chicken nuggets with respect to the different treatments during refrigerated storage are shown in p < 0.05) in regard to the coating formulations and storage time.The taste score values of chicken nuggets with respect to the different treatments during refrigerated storage are shown in The odor score values of the chicken nuggets during refrigerated storage are shown in p < 0.05) in regard to the coating formulations and storage time.The overall acceptability of uncoated chicken nuggets samples, SA (1.5%), PPP (1.5%), and SA (1.5%) + PPP (1.5%) during refrigerated storage are given in Different plant sources comprise a vital source of chemical compounds, including antimicrobials and antioxidants and total phenolic compounds. The peel of fruits contains these chemicals in abundant quantities. However, the pomegranate peel is a rich source of bioactive chemicals that play valuable role in the preservation of food products. In the current study sodium alginate and pomegranate peel powder were used separately and in combination as coating materials. Results showed the potential antimicrobial and antioxidant properties of the coating formulations used for the chicken nuggets. In another study, a natural antimicrobial and nontoxic substance called chitosan was used as a coating material, comprising however, a double practical component on the stability and shelf-life extension of foods. Chitosan plays also a significant role in the inhibition of fungus progression, by decreasing the fungus cell wall structure of protein inhibitors .Moosavi-Nasab et al.  carried Green tea extracts (rich in antioxidants and polyphenols) were used in food to prevent the oxidation process in triglycerides . Lipid o2 cells.GSE (grape seed extract) is labeled as a strong antiradical substance possessing scavenging actions beside free fundamental and Schiff-base metal complexes, as well as synergistic achievement with other antioxidants . YingyuaA previous study indicated that edible coating of cashews reduces lipid oxidation by protecting them from oxygen exposure during storage . During p < 0.05) were occurred in both coated and uncoated samples of chicken nuggets. The coated materials PPP and SA + PPP are vital sources of antioxidants that resulted in the lower POV in the coated chicken nuggets samples as compared to the uncoated ones. These results are in agreement with those of a previous study [The current results of the POV showed that significant differences . These findings are in agreement with the results reported in previous studies ,53. DuanDashti et al.  reportedPomegranate peel powder (PPP) and sodium alginate (SA) were used for the preparation of the edible coating of chicken nuggets. Physicochemical, microbiological, and antioxidant parameter analyses of the product were determined at storage intervals of 0th, 7th, 14th, and 21st day. Chicken meat was procured from local market of Faisalabad, Pakistan. Pomegranate peels were also collected from local markets and were dried. After the drying period, the dried peels were converted into powder and stored in air-tight containers for further analysis. All chemicals and reagents were purchased from Sigma Aldrich .Chicken meat nuggets were prepared following the method of Perlo . Firstly2) for 1 min. After the coating of nuggets, these were kept in a hot air oven at 40 °C for 30 min for the efficient casting of coating over the nuggets. The nuggets were coated with SA (1.5%), PPP (1.5%) and SA (1.5%) + PPP (1.5%) and packed properly, along with the uncoated nuggets which served as the control samples. These were labeled and kept at 4 ± 1 °C. This experimental product was evaluated at different storage time intervals , on the basis of physicochemical, sensory, and microbiological analyses.The experimental product was coated with 1.5% solution of SA, PPP and SA+ PPP Microbial load  of different treated chicken nuggets were determined according to the method described by Helrich . Chickeng) for 60 s. Then, 1 mL of the standardized solution was transferred to a test tube and the estimation of the lipid oxidation was determined by using the method of Ahn et al. [g) for 13–15 min and kept in room temperature. Spectrophotometer  was used to measure the absorbance of the nuggets at 532 nm. TBARS values were estimated by measuring by using the following equation:Chicken nugget samples (5 g in 15 mL of distilled water) were standardized , as described by Koniecko . Spectros = Absorbance of the sample, Ab = Absorbance of the blank, M (Standard) = 41.52, m0 = mass in gram, 55.84 = atomic weight of iron.A2CO3 (5%) was added and the mixture was vortexed. Then, the test tube was kept in a dark room for 1 h. Spectrophotometer  was used to measure the absorbance at 725 nm. Total phenolic content was expressed as gallic acid equivalents using a standard gallic acid curve (mg of GAE/g).Total phenolic content of chicken nuggets (uncoated and coated) was determined according to the method of Tezcan and Sever, . The samThe DPPH inhibition (antioxidant activity) of the chicken nuggets was estimated according to the method of Brand-Williams et al. . The chiThe pH of chicken nuggets samples was measured with a pH meter . Distilled water (50 mL) was mixed with chicken nuggets (10 g) and the pH-meter was immersed to the obtained solution to measure the pH value.The color of chicken nuggets samples was measured with a Hunter colorimeter  at storage intervals . The chicken nuggets were kept in a plate under the photocell. The color of chicken nugget samples surface was determined by the Hunter colorimeter with measurements standardized with regard to a white calibration plate . CIEb * (yellowness), CIEL * (lightness), and CIEa * (redness) are the average values of 8 random reads of different viewing apertures of the samples.Sensory evaluation of the different treated chicken nuggets samples was conducted on the basis of color, flavor, texture, taste, and overall acceptability following the procedure of Meilgaard et al. . The trap < 0.05) was determined by applying analysis of variance (ANOVA), following the principles outlined by Steel and Torrie [The obtained data were subjected to statistical analysis by applying a complete randomized design (CRD). Level of significance  and PPP (1.5%) chicken nuggets and lowest in the uncoated samples. The minimum values of total aerobic count and coliform were found in the combination of SA (1.5%) and PPP (1.5%). In the coated chicken nuggets, higher L*, a*, and b* values were found at the end of storage, whereas acceptable sensory parameter values  were also observed in the coated chicken nuggets. Therefore, edible coating consisted of SA (1.5%) and PPP (1.5%) may provide better quality and safety characteristics for chicken nuggets during refrigerated storage.


LOD and LOQ were 1.12 and 3.39 ng mL−1, respectively. The intra-day and inter-day precision and accuracy were 1.17 to 2.75% and 97.86 to 101.48%, respectively. The intrinsic clearance (CLint) was 2.71 mL min−1 kg−1 and the in vitro half-life (t1/2) was 48.80 min. A review of the literature revealed that there are no previous articles about the quantification of OTB in human plasma using LC-MS/MS or its metabolic stability assessment.Olmutinib  is an orally available third-generation epidermal growth factor receptor tyrosine kinase inhibitor (EGFR TKI). It was developed by Boehringer Ingelheim and Hanmi Pharmaceutical Co. Ltd for the cure of non-small cell lung cancer (NSCLC). In May 2016, OTB was approved in South Korea for the treatment of patients suffering from metastatic or locally advanced EGFR T790M mutation-positive NSCLC. A LC-MS/MS methodology was validated for OTB quantification in human plasma. An extended application for this validated LC-MS/MS is OTB metabolic stability evaluation. Chromatographic separation of OTB and ponatinib  was attained using a reversed phase with isocratic elution. The linearity of the developed LC-MS/MS method ranged from 5.00 to 500.00 ng mL Olmutinib  is an orally available third-generation epidermal growth factor receptor tyrosine kinase inhibitor (EGFR TKI). The FDA granted a breakthrough therapy designation for OTB in NSCLC.10 In May 2016, OTB was approved in South Korea under the trade name Olita™ for management of patients suffering from metastatic or local advanced EGFR T790M mutation-positive NSCLC.10OTB HM61713,  is an or11 The reported method is a gradient method with a pharmacokinetic application. These findings motivated us to develop a validated isocratic LC-MS/MS method for estimating OTB with high recovery in human plasma. Our current developed proposed procedure is an isocratic method with very high reproducible recovery  for OTB quantification in human plasma with applications to evaluate OTB metabolic stability. For the evaluation of OTB metabolic stability, two important parameters were computed. These two parameters are intrinsic clearance and in vitro half-life (t1/2) that could be used for calculating in vivo t1/2, hepatic clearance and bioavailability, which gives an idea about the investigated compound metabolism. If the tested compound is rapidly metabolized in the human body, it can possess low bioavailability.12A systematic literature review for OTB showed that there is one reported analytical method to quantify OTB in rat plasma, where the percent recovery of OTB in rat plasma was 85.8% to 95.5%.2.2.1.13–16 Maintenance of the rats was done following the Animal Care Center guidelines at the College of Pharmacy at King Saud University (KSU) in Saudi Arabia. The animal experiment protocol was approved by the Institutional Review Board at KSU. The Ethics Review Committee at KSU approved the experimental design that was utilized for animal study.All chemicals and reagents are listed in 2.2.All liquid chromatographic and mass spectrometric parameters were adjusted to achieve fast separation with high resolution. PNT was chosen as IS in OTB analysis as the same method of extraction was applied successfully to OTB and PNT  and the elution time of PNT was close to that of OTB with good resolution; thus, the method is fast with a short run time. Both PNT and OTB are TKIs and are not prescribed together for patients; thus, this method can be utilized for clinical applications such as pharmacokinetics or therapeutic drug monitoring (TDM) for patients under olmutinib treatment.−1) was used as a drying gas in the ESI source and high-purity nitrogen (50 psi) was used as a collision gas inside the collision cell of the mass spectrometer. The ESI temperature and capillary voltage were adjusted to 350 °C and 4000 V, respectively. The Mass Hunter software of Agilent was used to control the instruments and manage the data acquisition. Quantifying OTB was done using the multiple reaction monitoring (MRM) analyzer mode for the mass transitions (parent to product ion) from 487 → 432 and 487 → 402 for OTB and from 533 → 433 and 533 → 260 for PNT (IS) (Detection was conducted on a QqQ mass detector connected to an electrospray ionization source (ESI) that was operated with a positive mode. Low-purity nitrogen (11 L minPNT (IS) . The fraPNT (IS) .2.3.−1) was solubilized in DMSO. Afterwards, the stock solution was diluted ten times with the mobile phase to prepare working solution 1 (100.00 μg mL−1) that was further diluted ten times with the mobile phase to prepare working solution 2 (10.00 μg mL−1). PNT stock solution  was solubilized in DMSO. A working solution of PNT was made by diluting stock of PNT (IS) 50.00 times with the mobile phase to prepare working solution 3 (2.00 μg mL−1).OTB (1.00 mg mL2.4.−1) was mixed with certain volumes of human plasma matrix to generate fourteen calibration points: 5.00, 10.00, 15.00, 20.00, 30.00, 40.00, 50.00, 80.00, 100.00, 150.00, 200.00, 300.00, 400.00 and 500.00 ng mL−1. Three calibration standards 15.00 ng mL−1, 150.00 ng mL−1 and 400.00 ng mL−1 were selected as quality controls: low (LQC), medium (MQC) and high (HQC), respectively. Extraction was done using a protein precipitation technique by ACN. First, alkalinization of spiked plasma standards was conducted by adding 1 mL of NaOH/glycine buffer  followed by mixing for 30 seconds using a vortex. Second, two mL of ACN was added, followed by centrifugation at 14 000 rpm (12 min at 4 °C) to aid in protein precipitation.17 Supernatants were collected and then, filtration was done using a syringe filter (0.22 μm pore size). Fifty μL of PNT working solution 3 was added to 1 mL of the filtered samples and transferred to HPLC vials to be loaded into LC-MS/MS for analysis. Similarly, blank samples were prepared by utilizing the stated mobile phase in place of plasma to verify the absence of any interference from plasma components at RT of OTB and IS. A calibration curve was constructed for spiked human plasma by plotting the peak area ratio of OTB to PNT (y axis) against OTB standard nominal values (x axis). A linear regression equation was used to express the linearity of the established method. The slope, r2 and intercept values were calculated.OTB working solution 2  and limit of detection (LOD). All these parameters were computed for OTB.The validation parameters of the LC-MS/MS methodology that was developed to estimate OTB in human plasma were mentioned in detail earlier.2.6.−1 microsomal proteins in triplicate. All incubations were kept for 10 minutes to reach 37 °C in a water bath. Initiation of the metabolic pathway was performed by adding 1 mM NADPH in phosphate buffer (pH 7.4) that contained 3.30 mM MgCl2. Two mL ACN was added to terminate the metabolic pathway at certain time points: 0.00, 2.50, 5.00, 7.50, 10.00, 15.00, 20.00, 40.00, 60.00, 90.00 and 120.00 min. The same method of extraction as mentioned above was utilized for OTB extraction. The metabolic stability curve for OTB was constructed.The metabolic stability of OTB was studied by measuring the decrease in OTB concentration when it was incubated with RLMs. One μM OTB was incubated with 1.00 mg mL3.3.1.Optimizations of the chromatographic parameters including pH, mobile phase constituents and RRLC column were conducted. The pH of the aqueous part (0.1% formic acid) was adjusted to 3.2 as at pH values higher than this value, increase in elution time and peak tailing were observed. The ratio of mobile phase components (aqueous phase to ACN) was adjusted to 60% : 40% as more ACN led to overlapping of peaks with poor resolution and less ACN led to a delayed elution time. Various columns (Hilic columns) were checked but OTB and PNT were not retained. The best outcomes were obtained using a C18 column.The chromatographic separation of OTB and PNT (IS) was attained in 3 min. OTB and PNT chromatographic peaks resolved well in the absence of carryover in the blank samples: either the plasma matrix sample or plasma plus internal standard. 3.2.3.2.1.3.2.2.y = ax + b). A linear fit was verified by the coefficient of determination (r2), which was linear in the range from 5 ng mL−1 to 500 ng mL−1. The recovery for OTB detected in the spiked human plasma samples was 99.61 ± 2.82% with RSD less than 2.17%.The least square statistical method was utilized to compute calibration curve equations (r2) for the supposed methodology were 5–500 ng mL−1 and ≥0.9999, respectively, in human plasma. The calibration curve regression equation of OTB in human plasma was y = 3.69x − 5.67. LOD and LOQ were 1.12 and 3.39 ng mL−1, respectively, in human plasma.The linearity range and correlation coefficient  confirmed the performance of the developed methodology. The reproducibility and repeatability of the method were represented by the intra-day and inter-day accuracy and precision, which were 1.17 to 2.75% and 97.86 to 101.48%, respectively . The mea3.2.3.19As mentioned in 3.2.4.−1) and PNT (50 ng mL−1). The above-mentioned batches were labeled as set 1. Preparation of set 2 was conducted using a similar method, but the mobile phase was used instead of human plasma. Thus, the matrix effect was calculated utilizing the following equation:The studied human plasma that contains OTB and PNT showed values of 98.4 ± 2.5% and 98.3 ± 1.54%, respectively. The internal standard normalized matrix effect  was computed from the following equation:20 Accordingly, these results proved that the human plasma matrix has no noticeable effect on the ionization of OTB and PNT (IS).The IS normalized MF is 1.0 and it lies within the acceptable range.3.2.5.QC samples were used for stability studies of OTB under many conditions. 3.3.y-axis against time of incubation on the x-axis  on the e x-axis . The linThe slope was 0.0142.in vitro t1/2 method12 using the following equation:The intrinsic clearance of OTB was calculated following the int (2.71 mL min−1 kg−1) and a longer in vitro t1/2 value (48.8 min), which resulted in slower clearance of OTB from the blood by the liver and probable high in vivo bioavailability , eco-friendly , rapid (run time = 3 min.) and accurate (R% = 98.4 ± 2.5%). The adopted LC-MS/MS methodology was utilized for OTB metabolic stability assessment in RLM matrix using two terms: in vitro t1/2 (2.71 min) and intrinsic clearance (48.8 mL min−1 kg−1). From these two values, we concluded that OTB can be classified among low-extraction ratio drugs and hence can be slowly eliminated from the human body.A reliable and precise LC-MS/MS methodology was established for assaying the newly approved drug OTB in plasma. The established method is very sensitive (LOD = 1.12 ng mLMWA, AAK and HWD participated in putting the design of the research in addition to guiding the practical work. HWD, ASA and MWA performed the optimization and method validation experiments and manuscript writing. All the authors participated in revising and approving the final draft of the manuscript.The authors declare that they have no competing interests.CECollision energyEGFREpidermal growth factor receptorFVFragmentor voltageHQCHigh quality controlISInternal standardIS normalized MFInternal standard normalized matrix effectLC-MS/MSLiquid chromatography tandem mass spectrometryLODLimit of detectionLQCLower quality controlLOQLimit of quantificationMQCMedium quality controlNSCLCNon-small cell lung cancerOTBOlmutinibPNTPonatinibQCQuality controlTKITyrosine kinase inhibitor


CD33 rs12459419 CC genotype might benefit from the addition of GO to intensive treatment in contrast to patients with CT/TT genotypes. Nevertheless, contradictory results have been reported. We sought to shed light on the prediction of GO response in AML patients with rs12459419 polymorphism who were treated with GO in the consolidation (n = 70) or reinduction (n = 20) phase. The frequency distribution of the rs12459419 polymorphism in the complete cohort of patients was 44.4% (n = 40), 50% (n = 45), and 5.6% (n = 5) for CC, CT, and TT genotypes, respectively. Regarding the patients treated with GO for consolidation, we performed a Kaplan-Meier analysis of overall survival and relapse-free survival according to the rs12459419 polymorphism (CC vs. CT/TT patients) and genetic risk using the European Leukemia Net (ELN) 2010 risk score. We also carried out a Cox regression analysis for the prediction of overall survival, with age and ELN 2010 as covariates. We found no statistical significance in the univariate or multivariate analysis. Additionally, we performed a global Kaplan-Meier analysis for the patients treated with GO for reinduction and did not find significant differences; however, our cohort was too small to draw any conclusion from this analysis. The use of GO in consolidation treatment is included in the approval of the compound; however, evidence regarding its efficacy in this setting is lacking. Rs12459419 polymorphism could help in the selection of patients who might benefit from GO. Regrettably, in our cohort, the rs12459419 polymorphism does not seem to be an adequate tool for the selection of patients who might benefit from the addition of GO in consolidation cycles.Gemtuzumab ozogamicin (GO) is a conjugate of a monoclonal antibody and calicheamicin, which has been reapproved for the treatment of acute myeloid leukemia (AML). AML patients with the  Acute myeloid leukemia (AML) is a heterogeneous disorder characterized by a clonal expansion of myeloid progenitors . AML remGO treatment has been associated with hepatotoxicity and hepatic veno-occlusive disease/sinusoidal obstruction syndrome (VOD/SOS), myelosuppression, bleeding/thrombocytopenia, infusion-related reactions, and tumor lysis syndrome. In the ALFHA 701 study, drug-induced neutropenia was more prolonged in the GO group than in the control group after each cycle of consolidation. In addition, the median time to recovery of platelets was longer for patients in the GO arm than in the control arm for each treatment course (persistent grades III-IV thrombocytopenia was observed in 20% of patients in the GO group vs. 2% in the control group) .NPM1-mutated AML adult patients treated with GO during the induction phase suggested a potential value of CD33 genotype as a predictor of response [CD33 rs12459419 CC genotype (about 50% of study entrants) seemed to have a significantly lower risk of relapse and better event-free survival and disease-free survival after GO therapy, whereas this benefit was not seen in patients with the CT or TT genotypes. Rs12459419:c.41C > T; p.Ala14Val is related to the loss of exon 2 in the CD33 transcript, which results in a shorter isoform of the CD33 protein product (D2-CD33) that lacks the IgV domain. This domain, which is encoded by exon 2, is recognized by the antibody that is conjugated to calicheamicin in GO [Because GO is ineffective in many patients and/or may have adverse effects, there is interest in understanding favorable and unfavorable prognostic factors. Therefore, it is important to further refine the selection of patients that might achieve a favorable response from GO administration. Two studies conducted in pediatric and response , 13. Howresponse , 15. Spein in GO –19. ConsCD33 rs12459419 polymorphism influences the clinical outcome in adult AML patients treated with GO during consolidation or reinduction cycles.In the present study, we analyzed whether CD33 polymorphism determination in 90 patients: 48 men and 42 women. The median age was 55.3 years (range 17.4–76.9 years) in these patients with newly diagnosed AML between 2005 and 2013. Seventy patients received GO 3 mg/m (2) in the consolidation cycle and 20 in reinduction treatment. Finally, 19 patients underwent autologous hematopoietic stem-cell transplantation (HSCT), and 33 patients went through allogeneic HSCT.The PETHEMA AML epidemiologic registry (NCT02607059) includes the data of patients diagnosed with AML, regardless of the treatment administered. The primary patient and disease characteristics were collected retrospectively . We revin = 85) and RNA (n = 14) samples from seven centralized PETHEMA laboratories. The samples analyzed in this study were collected at the diagnostic stage. DNA was extracted using automated or manual DNA extraction kits  following the manufacturer's recommendations. DNA quantification was done using NanoDrop  or Qubit fluorometer  [CD33 SNP using HaeIII restriction enzyme digestion of polymerase chain reaction (PCR)-generated amplicons. The cDNA was synthesized using the High-Capacity cDNA Reverse Transcription kit . PCR products of 266 base pairs (bp) and 204 bp were generated from genomic DNA and cDNA, respectively, using HotStarTaq® DNA polymerase  using the manufacturer's recommended conditions and primers  at an annealing temperature of 66°C. Products were digested overnight with HaeIII  and separated on a 3% agarose gel to discriminate between the C allele (94 + 29 + 143 bp) and T allele (94 + 172 bp) using the DNA and C allele (61 + 143 bp) and T allele (204 bp) using the cDNA.We received DNA  . Samplesμl) and neutrophil recovery (>500/μl) among patients who received 3 + 7 plus GO as a consolidation cycle. A P value below 0.05 was considered statistically significant. SPSS version 19.0  was used for the analyses.OS was calculated from the date of diagnosis of AML until death in all included patients. RFS was calculated from the date of achieving complete response (CR) or CR with incomplete hematologic recovery (CRi) until the date of relapse or death due to any cause. CR and CRi were defined according to current 2017 ELN guidelines . OS and n = 90) were 44.4%, 50%, and 5.6% for the CC, CT, and TT polymorphism, respectively. From the total of 70 patients treated with GO during the consolidation cycle, there were 32 patients (45.7%) with CC genotype and 38 (54.3%) with CT/TT genotype. The median duration of OS in the CC group was 2.7 years  vs. 4.4 years (95% CI: 0.5-8.2) in the CT/TT group (P = 0.9). The median RFS in the CC group was 1.5 years (95% CI: 1.1-1.9) vs. 1.7 years (95% CI: 0.7-2.6) in the CT/TT group (P = 0.5).The frequency distribution of the rs12459419 genotypes in the whole patient population (P = 0.9). The OS in the intermediate group was 2.5 years (95% CI: 1.6-3.5) in CC genotype and 1.7 years (95% CI: NC-7.8) in CT/TT genotype (P = 0.9). The OS in the adverse group was 0.8 years (95% CI: NC-3.0) in CC genotype and 0.1 years (95% CI: 0.4-0.9) in CT/TT genotype (P = 0.8). The RFS in the favorable group was 1.7 years (95% CI: 1.3-2.7) in CC genotype and 2.7 years (95% CI: NC-7.5) in CT/TT genotype (P = 0.6). The RFS in the intermediate group was 1.4 years (95% CI: 1.3-1.6) in CC genotype and 1.6 years (95% CI: NC-9.4) in CT/TT genotype (P = 0.6). The RFS in the adverse group was 0.2 years (95% CI: NC-1.2) in CC genotype and 0.6 years (95% CI: 0.3-0.8) in CT/TT genotype (P = 0.4) . Rs12459419 genotype was not an independent predictor factor . Proportional hazard assumptions were checked by adding the interaction of every variable with time  . Cox regith time .P = 0.3). The distribution of rs12459419 genotype in RFS analysis was nine patients (45%) with CC genotype and three (15%) with CT/TT genotype. The median duration of RFS in the CC group was 0.6 years (95% CI: 0.3-0.9) vs. 0.6 years (95% CI: 0.5-2.7) in the CT/TT group (P = 0.8).  with CC genotype and eight (40%) with CT/TT genotype. The median duration of OS in the CC group was 0.6 years (95% CI: 0.5-0.8) vs. 0.3 years (95% CI: 0.3-0.3) in the CT/TT group (P = 0.3). In addition, we did not find statistical differences regarding the time to recovery of platelets: a median of 25.5 days (range 23-54) in the CT/TT group and 25.5 days (range 20-43) in the CC group (P = 0.8). No patient developed VOD/SOS during cycles of treatment prior to allogeneic HSCT. Regrettably, data on the incidence of VOD/SOS in the postallogeneic HSCT setting were not collected.The drug-induced neutropenia was similar in the CT/TT and CC genotype groups after the first cycle of consolidation with 3 + 7 plus GO: a median of 23 days (range 19-31) vs. 20 days (range 14-28), respectively  or consolidation treatment (n = 70) following the PETHEMA 2007 protocol. In our cohort, the rs12459419 genotype does not have a prognostic impact in terms of OS or RFS among AML patients treated with GO in either the consolidation or reinduction cycle. Additionally, the time to hematological recovery after the first consolidation cycle with 3 + 7 plus GO cycle was not influenced by the SNP genotype.Personalized medicine is based on selecting an adequate treatment for every individual patient . We evalCD33 rs12459419 polymorphism [NPM1 mutation showed improved RFS and reduced cumulative incidence of relapse in patients with CC genotype treated with GO during the induction phase [n = 113) did not reveal a significant impact of the SNP on OS or RFS. However, this study was performed in a patient population unlikely to benefit from GO as it included high-risk myelodysplastic syndrome patients and AML patients with unfavorable risk features [CD33 rs12459419 polymorphism. Regrettably, our study has shown that this polymorphism does not have prognostic value in this setting.The clinical impact of the genotype of this splicing polymorphism in AML patients treated with GO-containing chemotherapy is controversial. In a study in a mainly pediatric population, but also including young adults (COG-AAML0531) with AML (0-29 years old), a benefit of GO was demonstrated in the induction treatment in those with CC genotype of on phase . Converson phase . Moreovefeatures . In contfeatures . Neverthfeatures . Indeed,Our study has some limitations. First, our patients were selected from an observational registry, which can be interpreted as a limitation given the heterogeneity of treatment or as a strength because our data are more real-life than those derived from a clinical trial. Second, since the number of patients was small, it would be necessary to increase the sample size to validate our results. However, our results support the findings of the previously published study with the largest cohort , 15.In the event that the genotype of the rs12459419 polymorphism played a role in the mechanism of action of GO, it would modify the effect of the drug on leukemia and normal hematopoiesis only in patients with CC genotype. We would expect a longer time to recovery of peripheral blood counts in these patients without an increase in this time in patients with CT or TT genotypes. In our cohort, we did not find differences in the recovery of neutrophils and platelets based on the SNP genotype. These results were similar to those recently published by Short et al., which support the lack of prognostic value of the rs12459419 polymorphism found in our cohort [CD33 rs12459419 polymorphism, but no conclusions can be drawn due to the reduced amount of patients analyzed. In the consolidation setting, we analyzed 70 patients and did not observe differences between groups. Therefore, CD33 rs12459419 polymorphism does not seem to be an adequate tool to select patients who might benefit from the addition of GO in consolidation cycles.In summary, in our population of 20 AML patients treated with GO in reinduction, we found no prognostic significance of


The study reported here was conducted in the more temperate, rainier region of Western Province, Zambia and was designed to confirm the primary vector species in region and to estimate corresponding rates of feeding from prototype attractive sugar bait (ASB) Sarabi Bait Stations.Attractive targeted sugar bait (ATSB) stations are a promising new approach to malaria vector control that could compliment current tools by exploiting the natural sugar feeding behaviors of mosquitoes. Recent proof of concept work with a prototype ATSBThe product evaluated was the Sarabi v1.1.1 ASB station, which did not include insecticide but did include 0.8% uranine as a dye allowing for the detection, using UV fluorescence light microscopy, of mosquitoes that have acquired a sugar meal from the ASB. A two-phase, crossover study design was conducted in 10 village-based clusters in Western Province, Zambia. One study arm initially received 2 ASB stations per eligible structure while the other initially received 3. Primary mosquito sampling occurred via indoor and outdoor CDC Miniature UV Light Trap collection from March 01 through April 09, 2021 (Phase 1) and from April 19 to May 28, 2021 (Phase 2).Anopheles funestus s.l., which was the most abundant species group collected , had the highest sporozoite rate , and accounted for 94.3% (66/70) of all sporozoite positive specimens. Of those An. funestus specimens further identified to species, 97.2%  were An. funestus sensu stricto (s.s.). Anopheles gambiae s.l. (96.8% of which were Anopheles arabiensis) is a likely secondary vector and Anopheles squamosus may play a minor role in transmission. Overall, 21.6%  of An. funestus specimens and 10.4%  of An. gambiae specimens collected were positive for uranine, translating into an estimated daily feeding rate of 8.9% [7.7–9.9%] for An. funestus (inter-cluster range of 5.5% to 12.7%) and 3.9% [3.3–4.7%] for An. gambiae (inter-cluster range of 1.0–5.2%). Feeding rates were no different among mosquitoes collected indoors or outdoors, or among mosquitoes from clusters with 2 or 3 ASBs per eligible structure. Similarly, there were no correlations observed between feeding rates and the average number of ASB stations per hectare or with weekly rainfall amounts.The dominant vector in the study area is Anopheles funestus and An. gambiae vector populations in Western Province, Zambia readily fed from the prototype Sarabi v1.1.1 ASB sugar bait station. Observed feeding rates are in line with those thought to be required for ATSB stations to achieve reductions in malaria transmission when used in combination with conventional control methods (IRS or LLIN). These results supported the decision to implement a large-scale, epidemiological cluster randomized controlled trial of ATSB in Zambia, deploying 2 ATSB stations per eligible structure.The online version contains supplementary material available at 10.1186/s12936-023-04491-9. Vector control has contributed substantially to the global reduction in malaria burden that has been observed since 2000 . In sub-However, the sustained efficacy of currently available tools is threatened. Key challenges include insecticide resistance, which is expanding and intensifying in vector populations across Africa –10, and One promising new approach to malaria vector control is the use of attractive targeted sugar baits (ATSBs), which compliment current tools by exploiting the natural sugar feeding behaviors of mosquitoes –17. In c® Sarabi Bait Station  was designed to attract and kill malaria vectors. Early proof of concept work guiding product development was completed in Mali, including a large-scale entomological field trial of early prototype Sarabi Bait Stations against Anopheles gambiae sensu lato (s.l.) in the tropical savannah of Kayes Region that demonstrated high ATSB feeding rates and significant reductions in An. gambiae density, biting rates, and overall entomological inoculation rate (EIR) /[2a]). The model also calculated upper and lower estimates of daily feeding rates and assumed a natural daily mortality of 9.6%, no excess mortality caused by the uranine dye, a constant mosquito population, and an additional 9% mortality caused by standard vector control interventions (IRS and ITNs). When investigated mathematically, the proportion of mosquitoes positive for uranine as a function of feeding rate varied only slightly for a variable mosquito population, hence the assumption of a constant mosquito population for simplicity.The proportion of mosquitoes collected that were positive for uranine dye was converted to an estimated daily feeding rate using an Imperial College, London mathematical model expressed as a polynomial equation of the form F = auing rate . The coeTo calculate the number of clusters required to accurately estimate a mosquito feeding rate of 7.5% per night, with a targeted 5% half width of confidence interval, an R function was written) to implement the formula for “Confidence interval for one proportion in a Cluster Randomized Design” as implemented in PASS software.K) for a targeted proportion (of feeding) at a desired confidence interval width (precision) for a range of potential scenarios. The different terms of the formula are:K is the number of clusters requiredP is the population proportion of female Anopheles mosquitoes that are positive for uranine. This proportion is computed from the feeding rate using a second order polynomialbased on previous laboratory studies measuring the duration of detection of Uranine in mosquitoes.4.d is half the Width of the Confidence Interval (WCI) associated with the feeding rate, that is precision. The WCI was chosen such that the lower confidence limits were at or above the threshold feeding rate of 2.5%. Note that, the narrower the WCI, the more precise the estimate of feeding rate will be. Narrower WCI lead to greater cluster requirements.The term 5.rho (ρ), set at 10%.The ICC (intra cluster correlation) is 6.M is the cluster size. The cluster size is a function of mosquito availability  and sampling effort.7.C is the coefficient of variation of cluster size, computed as the ratio between the standard deviation of the cluster sizes to the mean of cluster size, set at 0.4.The formula calculates the required number of clusters , C = 0.4 and ICC = 0.1. Results indicated that a minimum of 7 clusters was required to estimate a daily feeding rate of 7.5% (± 5%), which could indicate a daily feeding rate of or above 2.5% (the lower limit) considered to be sufficient to lead to a 30% reduction in malaria incidence.The biological assumptions involved are as follows:Meteorological data was collected using a Davis Vantage Vue Wireless weather station positioned centrally at the field laboratory in Kaoma Town . During the study period, the onset of the dry season resulted in much cooler and dryer conditions. Weekly averages for maximum daily temperature, minimum daily temperature, and daily rainfall varied from 30.5 °C, 18.6 °C, and 23.6 mm during the first 3 weeks of the study to 28.4 °C, 8.9 °C, and 0.00 mm during the final 3 weeks.Data were maintained, cleaned, transformed, and summarized by descriptive statistics (including species compositions and abundance) using Microsoft Excel v2111  and Tableau Desktop v2020.4.11 . Sata/SE 14.2  and R and RStudio  were used for formal statistical analyses, including a linear regression analysis with robust standard errors clustered at the study cluster level to compare average vector ASB feeding rates and overall vector abundance by (1) indoor and outdoor trap locations and (2) by study arm. Additionally, Spearman’s ranked test was used to evaluate potential correlations between cluster-average feeding rates and crude ASB station density (the number of ASB stations per hectare within a cluster), weighted ASB station density (the number of ASB stations per occupied hectare within a cluster), as well as between overall weekly feeding rates and weekly rainfall amounts under various time-lag scenarios. Unless otherwise described, primary outcomes are presented as point estimates with 95% confidence intervals in parenthesis. For the linear regression analyses, P-values are from an F-test that calculates the difference between average feeding rates and estimates the probability that the actual difference equals zero. For Spearman’s rank tests, P-values are from a t-test that estimates the probability that the actual correlation coefficient equals zero.Anopheles spp. mosquitoes were collected over 14,859 collection-nights , followed by An. squamosus , Anopheles tchekedii , An. coustani s.l. , Anopheles maculipalpis , and An. gambiae s.l. . Other species encountered, each comprising less than 1% of the total and fewer than 1,000 specimens, included Anopheles brunnipes, Anopheles rufipes, Anopheles gibbinsi, Anopheles pretoriensis, Anopheles implexus, and Anopheles pharoensis. 5% of the Anopheles specimens collected  were not identified to species because of poor physical condition.During the entire 14 weeks of CDC LT surveillance , a total of 144,550 P. falciparum sporozoites in the head and thorax. PCR species identification confirmed a total of 2,150 An. funestus s.l. specimens: of these, 97.2%  were An. funestus sensu stricto (s.s.), 2.6% (56) were Anopheles parensis, and very few were identified as Anopheles rivulorum (3) and An. rivulorum-like. Of the 1,238 confirmed An. gambiae s.l. specimens, 96.8%  were Anopheles arabiensis, 1.8% (22) were An. gambiae s.s., and 1.4% (17) were Anopheles quadriannulatus. Anopheles coustani s.s. was the only species identified from the Coustani group (586 specimens).A subsample of specimens collected were screened for species confirmation, blood-meal hosts, and presence of An. funestus s.s., with an overall sporozoite rate of 3.16% and accounting for 94% of all sporozoite-positive mosquitoes. Anopheles arabiensis is likely an important secondary vector, and An. squamosus may play a minor role in transmission as well. The most common blood-meal source identified was human (44%), while cow (32%) and goat (23%) were also common and the human blood index  varied substantially by species  and 98.2% of An. gambiae .During the 12 weeks of mosquito surveillance following ASB station installation, there were 12,124 distinct CDC LT collections recorded. There were 42,587 An. funestus from 5,198 to 227 (Coefficient of variation [CV] = 0.73) and for An. gambiae from 200 to 15 (CV = 0.65). However, the patterns in cluster-level abundance variation for each species were very tightly correlated .The overall abundance of each vector species by cluster is shown in Fig. An. funestus was 1.24 (CI95 0.97–1.51), indicating a slight, but not statistically significant, tendency to collect more mosquitoes indoors than outdoors. For An. gambiae, this tendency to collect more indoors compared to outdoors was significant, with an indoor to outdoor ratio of 1.42 (Cl95 1.19–1.65).The indoor to outdoor collection ratio for An. funestus specimens collected and 10.4%  of An. gambiae specimens collected were positive for uranine in the abdomen. For both species, the proportion of uranine positive was higher in male mosquitoes, 31.2%  for An. funestus and 20.6% (7/34) for An. gambiae, than in female mosquitoes, 21.3%  for An. funestus and 10.2%  for An. gambiae. While the overall number of male mosquitoes collected was quite low, it is thought that high bait station feeding rates among males could potentially augment the effectiveness of an ATSB intervention. However, existing modeling ) compared to An. gambiae females , a significant difference of 7.4%  , a sigAn. funestus was 8.9% (7.7–9.9%), with an inter-cluster range of 5.5% (4.7–6.4%) to 12.7% (11.7–13.4%) and a cluster average of 8.6% (7.5–9.7%). Estimated daily feeding rates for An. gambiae were lower, with an overall aggregate feeding rate of 3.9% (3.3–4.7%), an inter-cluster range of 1.0% (0.8–1.2%) to 5.2% (4.4–6.1%), and a cluster average of 3.7% (3.1–4.4%).Using an Imperial College, London mathematical model expressed as a polynomial equation of the form described in the previous section, the proportion of each vector species sample that was positive for uranine dye was converted into estimated daily feeding rates Fig. . FollowiAn. funestus and 3.5% (3.0–4.2%) for An. gambiae  overall aggregate daily feeding for An. funestus was 8.9% (7.7–9.9%), with an inter-cluster range of 5.5% (4.7–6.4%) to 12.7% (11.7–13.4%) and a cluster average of 8.6% (7.5–9.7%). Estimated daily feeding rates for An. gambiae were lower, with an overall aggregate feeding rate of 3.9% (3.3–4.7%), an inter-cluster range of 1.0% (0.8–1.2%) to 5.2% (4.4–6.1%), and a cluster average of 3.7% (3.1–4.4%).Using an Imperial College, London mathematical model expressed as a polynomial equation of the form described in the previous section, the proportion of each vector species sample that was positive for uranine dye was converted into estimated daily feeding rates Fig. . FollowiAn. funestus and 3.5% (3.0–4.2%) for An. gambiae  overall aggregate daily feeding for Additional secondary objectives of this validation study included exploring whether ASB feeding rates varied in correlation to the density of ASB stations within a cluster (the number of ASB stations per hectare) or with rainfall (mm/week).An. funestus, there were no correlations observed between weekly rainfall and either mosquito abundance or ASB feeding rates in any of the scenarios. For An. gambiae, both outcomes were somewhat positively correlated with rainfall, with the strongest relationship observed with no time-lag .There were no statistically significant correlations observed between the average number of ASB stations per hectare and the daily feeding rates of either vector species , 716 (10.5%) reported having damage, with 503 of the 716 damaged stations also reported as having leaks. Of the 6,824 ASB stations visited at least once during the duration of the study (Phase 1 & 2), 542 reported having mold growth. An independent review of photographs taken by ASB field monitors during Phase 1 was conducted in June 2021 to validate reporting of damage and mold. During Phase 1, field monitors reported damage of 123 ASB stations, whereas the independent reviewer (looking at the photographs) reported damage of 34 ASB stations. During Phase 1, ASB field monitors reported leakage in 99 ASB stations, whereas the independent observer reported leakage in 55 ASB stations. During Phase 1, ASB field monitors reported mold on 77 ASB stations, whereas the independent observer reported mold on 306 ASB stations. Overall, the ASB station monitoring and replacement protocols maintained a high level of compliance among participating households, as more than 82% of all ASB stations deployed were found in use and collected at the end of the study. The majority of those ASB station not collected were reported to have been removed and discarded by the households.An. funestus s.l., readily fed from the prototype Sarabi v1.1.1 ASB station with an overall daily feeding rate of around 8.9% (CI95% 7.7–9.9%). Anopheles arabiensis, a likely secondary vector in the study area, also fed from the ASB stations, albeit at a lower daily feeding rate of around 3.9%. (CI95% 3.3–4.7%) These estimated daily feeding rates exceed the target of 2.5%, which is associated by modeling with a potential minimum 30% reduction in malaria incidence, by a statistically significant margin.This ASB feeding validation study is part of a package of preliminary studies designed to validate the ATSB approach to vector control prior to the start of a larger, 2-year cluster randomized control trial with an active malaria case incidence cohort. The results presented here show that the dominant vector species in the study area, Results also show that while ASB feeding rates varied substantially from cluster to cluster, they were consistent among mosquitoes sampled indoors and outdoors. Furthermore, similar feeding rates were observed with deployment of two versus three ASB stations per eligible structure, and rates were similar across the range of ASB spatial densities (from 3.07 to 11.16 ASB stations per hectare) observed across the 10 study clusters.An. funestus and An. arabiensis were initially attracted to occupied houses within the study clusters in search of a blood meal, and that if unsuccessful in acquiring a blood meal they were successfully diverted to the ASB stations as an alternative. It could also be possible that host-seeking mosquitoes took a sugar meal prior to acquiring a blood meal, perhaps to replenish energy stores before completing the blood feeding process. Interestingly, mosquitoes that successfully fed on an ASB station were equally likely to have been sampled indoors or outdoors, indicating that these vector populations are predominantly opportunistic in the relative location of their blood-seeking, sugar-feeding, and resting locations as defined here—either indoors or immediately outdoors in the peri-domestic area. This could suggest potential for a well-balanced effect for similarly deployed ATSBs on the dominant vector species in western Zambia, complimenting the core vector control interventions to provide a substantial incremental effect.The high proportion of females in the samples caught in and around households, and the relative consistency in sugar feeding rates and high human blood meal indices observed within each cluster throughout this 12-week validation study, suggest that An. gambiae s.l. (likely predominantly Anopheles coluzzii) during similar studies in Mali [An. funestus, are within ranges that are expected to produce significant excess mosquito mortality, above that induced by standard vector control measures alone, to a degree that is likely to also produce significant reductions in malaria transmission [While the ASB feeding rates observed here are somewhat lower than the feeding rates reported recently for  in Mali , 20, it smission .This study had several limitations. A single UV fluorescence microscope for dye identification in mosquitoes was not sufficient to quickly handle the volume of mosquito samples that were collected and hence lengthening the time required for dye screening. This is in combination with some uncertainty around dye persistence in mosquito abdomens and whether this is consistent in laboratory and field conditions or across vector species, all of which could have implications on the modelled conversions between dye positivity rates and daily feeding rates. Another operational challenge encountered was the need to continually update the ASB monitoring forms and procedures throughout the duration of the study: overall, ASB field monitors were likely over-reporting damage to bait stations and under-reporting mold growth. Lastly, the COVID-19 mitigation steps required throughout, complicated the logistics for mosquito collections, bait station monitoring, and the breadth and depth of community engagement and sensitization meetings in the study areas. Strengths of the study included utilizing robust power calculations and a substantial sampling effort, including equal sampling indoors and outdoors in the event that sugar feeding varied by mosquito populations. Despite the additional COVID-19 complications, community leadership was continually engaged and interested in the study itself and its implications for further research and development of the ATSB paradigm, and community buy-in was generally good.An. funestus and An. gambiae vector populations in Western Province, Zambia each readily fed from the prototype Sarabi v1.1.1 ASB sugar bait station. Furthermore, the observed feeding rates are in line with those required for ATSB stations to achieve sufficient reductions in the proportion of older mosquitoes to lead to significant reductions in malaria transmission, when used in combination with conventional control methods (IRS or LLIN), compared to the use of conventional methods alone. As such, these results were used to support the decision to fully implement the ATSB cluster randomized controlled trial with epidemiological primary outcomes in Zambia, deploying 2 ATSB stations per eligible structure within each intervention cluster.This ASB feeding validation study demonstrated that Additional file 1: Table S1. Total number of mosquitoes collected during the trial. Morphological IDs are presented. Figure S1. Comparisons of the daily feeding rates calculated from the proportions of mosquitoes collected that were dye positive, based on (a) the base-case scenario for An. funestus; (b) the alternate-case scenario for An. funestus; (c) the base-case scenario for An. gambiae; and (d) the alternate-case scenario for An. gambiae. Figure S2. No association between ASB feeding rates ASB spatial density, either crude numbers of ASB stations per hectare or weighted average ASB stations per occupied hectare, for (a) An. funestus or (b) weighted numbers of ASBs per hectare. Figure S3. Correlations between weekly rainfall amounts and (a) An. funestus feeding rates (b) An. funestus abundance (c) An. gambiae feeding rates, and (d) An. gambiae abundance. Figure S4. Cumulative dye positivity by study arm, post-crossover.


Rheumatoid arthritis (RA) is commonly associated with higher rates of comorbidities. Recent recommendations highlight screening comorbidities during the disease course because of their impact on patients’ ability to function, on disease outcome, but also on treatment choices. Hence the interest of our study that aimed to quantify the impact of comorbidities among RA patients using a validated tool the Rheumatic Disease Comorbidity Index (RDCI) and to explore the association between comorbidities and disease characteristics.We conducted a cross-sectional study over 12 months period, including patients followed for an established RA according to the ACR/EULAR 2010 criteria and hospitalized in our rheumatology department. Patients’ characteristics and disease features were collected for each patient. Comorbidities were quantified using the RDCI. We looked for the association between RDCI and patients characteristics and RA parameters. Univariable and multivariable analysis were made.They were 280 patients: 233 female (83.2%) and 47 male (16.8%) with a mean age of 58.07 (SD 11.12) years. The mean follow-up period was 14.74 (SD 1.63) years. Comorbidities were noted in 133 patients (47.5%). The mean comorbidity score measured by the RDCI was 1.05 (SD 1.23). RDCI was positively correlated with age . RA patients whose age of disease onset exceeds 40 years have significantly higher RDCI (1.8 (SD 1.3) [CI 95%: 1.36–1.88] vs. 1.5 (SD 1.2), p = 0.007). Moreover, RDCI was significantly associated with the presence pulmonary involvement (p < 0.001) and ocular involvement (p = 0.002). RDCI was also associated with erosive RA (p = 0.006), the presence of atlanto-axial dislocation (p = 0.014), and coxitis (p = 0.029). Regarding therapy regimen, RDCI was statistically increased in patients receiving bDMARDs compared to patients under csDMARDs (2.8 (SD 1.6) vs. 1.0 (SD 1.0), p = 0.021).In this study, comorbidity index was associated with signs of poor prognosis such as erosions, coxitis, and atlanto-axial dislocation. This confirmed the hypothesis that comorbidity can be a threat to the improvement in the long-term prognosis in RA patients. Patients with rheumatoid arthritis have higher rate of comorbidity which may impair their quality of life and affects disease outcomes.Rheumatic Disease Comorbidity Index (RDCI) is an excellent tool to predict extra-articular manifestations and signs of poor prognosis such as erosions, coxitis, and atlanto-axial dislocation.An early screening and an adequate management of comorbidities may result in better disease outcomes. Rheumatoid arthritis (RA) is a chronic inflammatory and autoimmune disease that affects about 1% of the population worldwide . The eviFew authors were interested in studying the impact of comorbidities on the RA outcomes. Comorbidities influence differently RA outcomes such as mortality, hospitalization, and disability . In factA number of comorbidity instruments have been created to assess comorbidities in patients with rheumatic disease . ComorbiWe conducted a cross sectional study including patients followed for an established RA according to the American College of Rheumatology/European League Against Rheumatism (ACR/EULAR) 2010 criteria  and hospDemographic features  were recorded. The following disease characteristics were collected for each patient: disease duration, age at onset of RA, immunological profile for rheumatoid factor (RF) and for anti-citrullinated protein antibodies (ACPA), disease activity measured by the disease activity score 28 (DAS28) , functioComorbidities were quantified using the RDCI, which is a validated method to quantify comorbidities in RA . It compWe used SPSS software version 23.0 for statistical analysis. The categorical data were expressed as numbers and percentages and the continuous data as means and standard deviations. Data were normally distributed using the test Kolmogorov-Smirnov. We looked for the association between RDCI and patients characteristics and RA parameters. The Pearson’s χ2 test was used for studying the association between two independent qualitative variables. The correlation between two continuous variables was performed using the Pearson test and between two categorical variables using the Chi-squared test. The Student’s t-test was used to study the association between a categorical and a continuous variable. Welch’s Test was used for unequal variances. It was only applicable for atlanto-axial dislocation. Missing variables were addressed using imputation.A multivariable analysis by means of multiple linear regression were performed to investigate which variables were independently associated with RDCI. This analysis included variables that were considered significant with a p-value of 0.2 or less in univariable analysis. The p-value was considered statistically significant when the value was less than 0.05.The RA cohort consisted of 280 patients. They were 233 female (F) (83.2%) and 47 male (M) 16.8%) with a sex ratio M/F at 0.2. Table 6.8% withRegarding the disease activity, the mean DAS28 (CRP) was 4.76 (SD 1.19) with extremes varying from 2.05 to 7.47. The distribution of patients according to their disease activity was as follows: 11 (3.7%) in remission, 25 (9.0%) with low disease activity, 115 (41.4%) with moderate activity, and 129 (45.9%) with high disease activity. The mean HAQ was 1.4 (SD 0.5) with extremes ranging from 0.3 to 2.2. Among the RA cohort, 136 patients (63.0%) presented extra-articular manifestations: pulmonary involvement  ), nodular lung disease (n = 29 (24.6%)), usual interstitial pneumonia (n = 21 (17.3%)), bronchiolitis obliterans organizing pneumonia (n = 9 (7.2%)), pleural nodules (n = 9 (7.2%))), rheumatoid nodules , ocular involvement , pericarditis , kidney involvement  (4 cases of glomerulonephritis and one case of amyloidosis), and systemic vasculitis .Pelvic radiographs showed coxitis in 34 patients (12.1%). Coxitis was unilateral in 32 patients and bilateral in 2 patients. During the disease course, 43 patients (15.4%) developed atlanto-axial dislocation. Regarding therapy, the use of corticosteroid was noted in 245 patients (87.7%). csDMARDS were prescribed as follows: Methotrexate (n = 246 (87.8%)), Sulfasalazine (n = 32 (11.5%)), Leflunomide (n = 5 (1.8%)), and Hydroxychloroquine (n = 8 (3.0%)). bDMARDs were prescribed for 72 patients (25.7%).Comorbidities were noted in 133 patients (47.5%). Multimorbidity was identified at 60 patients 21.4%). The distribution of patients according to their different comorbidities is summarized in Table 1.4%. TheRDCI was positively correlated with age . It was significantly increased in patients aged 65 years or over (2.1 (SD 1.4) [CI 95%:1.56–2.4] vs. 1.5 (SD 1.2), p = 0.009). RCDI has a tendency to be different in terms of gender (F: 1.5 (SD 1.2) vs. M: 1.8 (SD 1.3), p = 0.058). Regarding disease characteristics, RDCI was correlated with age at disease onset . It was higher in patients whose age of disease onset exceeds 40 years (1.8 (SD 1.3) [CI 95%: 1.36–1.88] vs. 1.5 (SD 1.2), p = 0.007). RDCI was not correlated to the disease duration . We did not find a difference in RDCI according to the immunological profile (seropositive: 1.6 (SD 1.2) vs. seronegative: 1.4 (SD 1.1), p = 0.968) and (immunopositive: 1.6 (SD 1.2) vs. immunonegative: 1.4 (SD 1.2), p = 0.887). RDCI was not correlated neither with the DAS28 (CRP)  nor with the HAQ .The association between RDCI and main disease characteristics is summarized in Table Regarding therapy regimen, RDCI was statistically higher in patients treated with bDMARDS versus patients receiving csDMARDS (p = 0.021).).On multivariable analysis, associated factors to the RDCI were: the presence of pulmonary involvement  and the presence of ocular involvement   with a high percentage of multimorbidity. Comorbidities may precede or appear during the course of the disease. Some studies noted that the burden of comorbidity is common before and after RA diagnosis, and the rate of accumulation accelerates after RA diagnosis , 22. OurComorbidities may influence patients’ disability as well as frequency of hospitalization and mortality . In factIn the present study we looked for the association between RDCI and RA characteristics.In concordance with previous literature data, we found that RDCI was positively correlated with age and age at disease onset , 23, 24.Other factors related to the rheumatic disease may interfere and vary with comorbidities. Tidblad et al. found that the serological status may influence on patients’ comorbidities . SeroposMoreover, extra-articular manifestations in RA have risen with an increased mortality rate in severe forms . Their oIn our study, comorbidity index was significantly higher in the presence of extra-articular manifestation such as pulmonary and ocular involvement. In fact, pulmonary involvement in RA and comorbidities may share common pathogenic mechanism. Some diseases, such as cardiovascular pathologies and diabetes that participate in ageing phenomenon of the body, are associated with telomeres shortening . TelomerIn addition to that, comorbidities have been reported to be associated with functional disability and deterioration of quality of life of RA patients , 35. ButAs mentioned above in our results, comorbidities were not associated with the disease duration, the disease activity or with the functional impairment. This may be explained by the moderate average of the DAS28 and the HAQ. In the other hand, comorbidities were associated with the use of biologic drugs. In the literature, multimorbidity rise up the complexity of the disease management and it was shown to affect negatively therapeutic target within one year after initiating DMARDs . AlthougThe main limitation of the present study is the selection bias since we included hospitalized RA patients with more severe disease. Thus, these results may not be generalizable to other populations. Other greater studies are needed to approve our findings.In this study, we elaborated important findings expanding our current knowledge and enriched the available literature data. It seems clear that elderly patients face more comorbidities. Based on our results, we confirmed the association of comorbidities with severe outcomes and signs of poor prognosis such as structural damages for patients with RA. With increasing life-expectancy and improved control of joint disease in RA, the relative importance of comorbidity in RA is increasing and can be a threat to the improvement in the long-term prognosis in RA patients. Thus, we highlight the importance of managing comorbidities in this population as part of managing RA.