Solanum lycopersicum L.) is an annual vegetable cultivated all over the world. It faces biotic and abiotic stresses, such as salinity, in arid and semiarid regions. Investigating the relationship between physiological and economic traits, such as fruit yield, under stress conditions is necessary to identify tolerant genotypes. This study was conducted to identify tolerant tomato families according to the relationship between several important physiological, morphological and phenological traits. Twenty S3 families were cultivated in a factorial experiment  based on a randomized complete block design with three replications in 2019. Twenty physiological, agronomic and fruit-quality-related traits were investigated. Analysis of variance was used to prove the existing effective genetic diversity. Genetic diversity and the relationships between traits were graphically shown using heatmap clustering. Finally, genetic parameters, such as Pearson’s correlation, trait stability index and heritability were used to calculate the mathematical value of families using the Modified Analytical Hierarchy Process. Families exhibited different behaviours under normal and stress conditions. The tolerant families responded physiologically to the salt stress. Therefore, they reduced both cell membrane degradation and photosynthesis disruption by increasing proline, lycopene, carotenoid and sugar content. Therefore, fewer reductions in morphological traits were observed in these families. The most important traits based on the selection strategy were lycopene content, K+/Na+ ratio, days to flowering and biological yield. In addition, three families, H4/T/30/1, H1/T/12/5 and H1/T/47/4, were selected as the most suitable alternatives to construct the breeding population of the next generation.The tomato ( Investigating the relationship between physiological and economic traits, such as fruit yield, under stress conditions is necessary to identify tolerant genotypes. This study was conducted to identify tolerant tomato families according to the relationship between several important physiological, morphological and phenological traits. Finally, genetic parameters, such as Pearson's correlation, trait stability index and heritability, were used to calculate the mathematical value of investigated families using Modified Analytical Hierarchy Process. Families exhibited different behaviors under normal and stress conditions. The tolerant families responded physiologically to the salt stress. Therefore, they reduced both cell membrane degradation and photosynthesis disruption by increasing proline, lycopene, carotenoid and sugar content. Fewer reductions in morphological traits were observed in these families. The most important traits based on selection strategy were lycopene content, K+/Na+ ratio, days to flowering and biological yield. The tomato is an annual vegetable that belongs to the Solanaceae family. It is consumed in several ways, including fresh, cooked and canned. It has also been used in processed products, such as juice, pulp, paste and several types of sauces. . Salinit+/Na+ ratio  using multivariate statistical methods.−1) and the second level was irrigation with salt water (EC = 12 dS m−1), according to −1.Twenty-one S3 families were obtained from a breeding program with four generations of self-pollinated tomato hybrids from 2015 to 2018 . The exp−11; pH = 6.0), which was replaced with irrigation. Sandy loam soil  consisting of 700 and 125 mg kg−1 K and P as well as 0.12 % N, 1.21 % organic carbon and 21.1 % fixed carbon was used. It should be noted that, according to the specific soil texture used in this experiment, due to the different behaviours of different soil types regarding salinity, the results may differ in clay-textured soils.Since the planting medium was in pots and the controlled glasshouse conditions were considered for plant growth, there was no difference between direct sowing and indirect culture (using plantlets and transplanting) concerning the germination rate, and due to the greater uniformity of plants in direct sowing, this method was performed. Before sowing, the seeds were sterilized with sodium hypochlorite solution (10 %) for 5 min and then in ethanol (96 %) for 1 min and thoroughly washed with sterile distilled water. Seeds were sown in plastic pots (30 cm diameter and 40 cm height) filled with Hoagland’s solution  and Ca2+/Na+ ratio (Ca/Na) were investigated.Twenty physiological, agronomical and fruit quality-related traits, including days to 50 % flowering (DTF), days to 50 % fruit formation (DFF), plant height (H), biological yield (BY), fruit number (FN), stem diameter (SD), fruit yield per plant (Yld), total soluble solids (TSS), lycopene content (Lyc), salinity , pH, sugar content (Sug), electrolyte leakage (EL), relative water content (RWC), proline content (Prol), relative chlorophyll content (SPAD), the efficiency of photosystem II (Fv/Fm), carotenoid (CAR), KMorphological traits were measured at the end of the harvest in each plant. The average of samples’ data in each replication was recorded. Picking fruits and assessing their characteristics were done in the full ripening stage  based on 2 leaf discs were provided from the bulked sample and submerged in 10 mL of deionized water in 20-mL screw-cap vials and kept at 25 °C in the dark overnight. The electrical conductance of the solutions was measured with a conductivity meter . Then, the samples were autoclaved and total electrical conductance was quantified after vigorously shaking the vials. All measurements were recorded at 25 °C. Electrolyte leakage of samples was estimated by To measure EL according to C1 and C2 are the electrical conductivity of the sample solutions before and after autoclaving, respectively.where To calculate RWC, fresh weight (FW), dry weight (DW) and turgid weight (TW) of the samples were determined and RWC was calculated by −1) was determined according to the protocol used by Proline content per fresh weight  in the presence of the LaCs reagent , 536.9 is the molecular weight of lycopene (g mol–1), V is the volume of the nonpolar phase (mL), W is sample weight (kg) and 17.2 is the molar extinction coefficient for lycopene in hexane (L mol–1 cm–1).where Tomato fruit juice was provided to use by a digital refractometer to measure TSS and express results in Brix . Also, the pH value of the tomato juice was determined using a pH meter . The salinity of the tomato fruit juice was determined with an electrolytic conductivity meter .After investigating assumptions of ANOVA, a combined ANOVA with a linear model based on RCBD was done by the model:where Pearson correlation between all traits with fruit yield, fruit lycopene content and DFF was calculated.The stability of traits in different conditions was calculated using regression analysis between trait data in two experiments, where data in normal conditions were considered as independent variables and data in stress conditions as dependent variables. The absolute regression coefficient showed the stability of the trait. The lower the value of the absolute regression coefficient, the higher the trait stability. In this paper, the index calculated by this method is called stability index (SI).seeCluster analysis of traits and genotype × experiment was performed, and total variation was simultaneously categorized by heatmap graphical analysis. AHP with some modifications (MAHP) according to Step (1) Aligning traits and families .Step (2) Calculating the weight of traits.Step (3) Calculating the weight of families.ANOVA, stability index, and MAHP calculations were done by +/Na+ ratio. It revealed the uniformity of the glasshouse environment and the appropriate control of the conditions. The calculated coefficient of Determination (R2) for almost all traits was more than 0.7. The minimum value of R2 (0.33) was found in pH. As a result, the salinity effect on pH was not statistically significant.seeAccording to the different families’ reactions to salinity stress and the significant interaction between genotype and environment, treatment combinations were compared using Duncan’s multiple range test and the related table is available on the journal’s website see. A wide In general, the families with more changes in physiological manners showed fewer changes in their performance. For example, salt stress had the highest effect on families 12 and 4 which did not show significant changes in physiological traits. Also, the lowest effect of salinity stress was on the performance of families 2 and 16, which showed significant changes in the biosynthesis of proline, sugar and carotenoid. Families 2, 8 and 9 showed the best performance under salt conditions. Also, changes in fruit yield were correlated with the fruit number under stress conditions while being correlated with the stem diameter and biological yield changes under normal conditions. The best families based on early ripening, fruit yield and fruit lycopene content under normal conditions were 12, 4 and 2.According to +/Na+ ratio, Ca2+/Na+ ratio, stem diameter, fruit yield, plant height and biological yield and the second one included fruit number, relative water content, relative chlorophyll content and efficiency of photosystem II. All traits in both groups were entirely affected by stress. Measured proline, sugar, carotenoid, lycopene and salinity were classified in the third group, which increased under salinity stress. The fourth group included pH, total soluble solids and electrolyte leakage, which did not show a recognizable pattern in the two experiments. In other words, the effect of salinity stress on the last group of traits was not impressive compared to the genotype’s effect. Therefore, the only effective factor in creating diversity regarding the third group of traits is the genotype.Traits were classified into four groups and families were classified into two groups in each experiment. The first group consisted of days to ripening, KIn normal conditions, families 3, 4, 5, 6, 9, 11, 12, 13, 16 and 18 were in a group with the highest values of morphological traits, such as yield and yield components. Another group of normal conditions consisted of families 1, 2, 7, 8, 10, 14, 15, 17, 19 and 20 with lower values of morphological traits. The two mentioned groups were similar in physiological and phenological traits under normal conditions. Families 2, 4, 5 and 12 with high amounts of yield and yield components and low amounts of phenological traits exhibited suitable agronomical characteristics under normal conditions. Considering stress conditions, families 1, 3, 5, 6, 8, 9, 10, 15, 16, 17 and 19 were in the group with the highest amounts of morphological and physiological traits. Another group included families 2, 4, 7, 11, 12, 13, 14, 17, 18 and 20 with higher values of phenological traits. The more stress-tolerant families were categorized in the first group of stress conditions. High tolerance to salinity stress in this group was due to their physiological responses, such as increased biosynthesis of proline, sugar, lycopene and carotenoid resulting in increased cell membrane resistance, increased photosynthesis and finally a better plant phenotype and fruit yield.+/Na+ ratio, ripening and biological yield were the most important traits in the selection, respectively. On the other hand, electrolyte leakage, pH, total soluble solids, relative water content and sugar content of fruit juice had the lowest effect on selection, respectively.According to The importance of traits was assessed based on their total effects on target traits, including fruit yield, days to ripening and fruit lycopene content, determined by Pierson’s correlation. The stability index introduced in this study was another parameter that had an impact on their importance. In other words, a multivariate selection was done for multiple purposes.Based on these weights, the families were sorted using a capability index . FamilieThese families were determined as tolerant by heatmap, GE-clustering and MAHP as the best families for breeding to make the next generation population and gain premature salt-tolerant families with high fruit yield and high fruit quality.Comparing different characteristics of a single genotype as well as comparing differences between families regarding a single trait can be exhibited using a heatmap . They were more likely to produce suitable progenies than other families based on the breeding strategy.+/Na+ ratio, days to flowering and biological yield. Families of 21, 7 and 20 were identified as the best choices for breeding and creating the next-generation population. It was explained that selecting these families may lead to increasing the possibility of achieving premature salt-tolerant families with high fruit yield and quality.Salinity stress had significant effects on investigated families. ANOVA table, heatmap and GE-clustering revealed the different manners of families in face of the salt stress. Morphological characteristics and especially traits related to fruit yield were severely reduced under salt stress. The decreased yield of stress-tolerant families was less than others due to the appropriate physiological responses. Tolerant families reduced degradation of the cell membrane and disruption of photosynthesis by increasing biosynthesis of proline, lycopene, carotenoids and sugar, under salinity stress to reduce the effect of salinity on plant vigour. Stress-tolerant families were distinguished from sensitive families by different multivariate methods as families 16, 8 and 1. The most important traits for breeding strategy were identified by MAHP as lycopene content, KThe following additional information is available in the online version of this article –MAHP. MATLAB script for calculating MAHP.Duncan’s Multiple Range Test.plad037_suppl_Supplementary_MaterialClick here for additional data file.plad037_suppl_Supplementary_Material_1Click here for additional data file.


Background: The incidence of cutaneous squamous cell carcinoma (cSCC) is rapidly increasing, paralleling the aging of the population. cSCC predominantly affects chronically sun-exposed areas, such as the head and neck region. At our tertiary center, a multidisciplinary approach to non-melanoma skin cancer is provided for locally advanced cSCC. Methods: We retrospectively revised all patients with locally advanced/metastatic cSCC treated with anti-PD1 antibody (Cemiplimab) at our Institution from January 2020 to March 2023 (minimum follow-up of 4 months on treatment). Results: Overall, we consecutively treated 20 ultra-octogenarian patients, of whom 15 were males and 5 were females (median age: 86.9 years). Despite age, a median number of concomitant drugs, and comorbidities, efficacy, and safety were superimposable with the available literature. No patients reported treatment-related adverse events of grade 3 or higher. Grade 2 adverse events were reported in 25% of patients. Overall, the response rate was 65%, with 50% partial responses and 20% long-lasting stable disease. The median duration of response was 14 months. The G8 elderly score was assessed in all patients, and the median score was 12 (range 9–14). Conclusions: Among ultra-octogenarian patients, a clinical benefit from Cemiplimab was obtained in most, including tumor shrinkage and pain relief. Cemiplimab confirmed its effectiveness in elderly patients in a real-life setting, with no new safety concerns. Cutaneous squamous cell carcinoma (cSCC) is the second most common skin cancer, and its incidence is rapidly increasing in parallel with the aging of the population .Common risk factors for cSCC are older age, immunosuppression, and chronic sun exposure. Surgery is the mainstay for early-stage  disease, while locally advanced disease requires a multidisciplinary assessment .Immunosuppression is strongly associated with an increased risk of developing cSCC. Transplant recipients have a 65–250-fold increase in risk of cSCC, and a 10-fold increase in risk. ,4,5 The    ,5,6,7,8. weight)  9,10]. ,10.   for the management of patients with skin cancer, and related challenges.We conducted a retrospective observational study to assess real-world treatment compliance and outcomes in ultra-octogenarian patients with unresectable LA-cSCC or mcSCC. We used the electronic health records of patients who were treated with Cemiplimab from January 2020 to May 2023.The inclusion criteria were the following: age ≥ 80 years at the time of systemic treatment initiation; systemic treatment with Cemiplimab (at least 2 doses); and informed consent to data collection processing and privacy. The exclusion criteria were the following: age < 80 years; major contraindications to antiPD1 therapy; and patients’ refusal of Cemiplimab treatment.At our Institution, patients with locally advanced, recurrent, or metastatic cSCC are all evaluated by a skin cancer multidisciplinary team (MDT). The members of the skin MDT are dermatologists, radiation oncologists, medical oncologists, maxillo-facial surgeons, plastic surgeons, and pathologists. On-demand geriatricians and nutritionists are likewise involved.The Geriatric 8 (G8) questionnaire was administered prior to the start of treatment . We perfAdverse events (AEs) intensity was graded according to the Common Terminology Criteria for Adverse Events (CTCAE) version 5.0.Data collection was performed under normative regulations, indications, and restrictions on the matter of retrospective clinical studies.All patients provided written informed consent according to the actual rules for data collecting, processing, and privacy required in Italy.Overall, 20 patients who fulfilled the inclusion criteria were consecutively treated and enrolled in the present study. The main patient characteristics were the following: 15/5 M/F; a median age at treatment start of 86.9 years (range 80–103); and a median number of surgeries of 2 (range 0–4). Fourteen patients (79%) received prior radiotherapy for the primary cSCC after first diagnosis or at the time of disease recurrence.Overall, the G8 median score was 12 (range 9–14). Eight (40%) patients were assessed as vulnerable by the G8 score. These patients were evaluated by the MDT and referred for CGA. However, 6 out of 8 patients with G8 scores ≤ 14 did not receive a CGA because at that time, an onco-geriatric pathway was not available at our Institution.All six vulnerable patients who did not undergo CGA received at least two cycles of Cemiplimab. The median number of cycles at the time of the present analysis was 14.We tried to correlate the G8 score and the FRAIL score, however while the first allowed us to personalize the geriatric risk , the FRAIL score with a small range of 1–5 and a median value of 2 was used to indicate a yes or no condition.Seven patients (35%) suffered from metabolic syndrome, eleven patients (55%) had cardiovascular disease history, and five patients (25%) had chronic pulmonary disease. Two patients received Cemiplimab while on immunosuppressant therapy for hematologic disease.The median number of concomitant drugs was three (range 2–8); the majority included anti-hypertensive  and cholesterol-lowering drugs (statins).An objective response was observed in 18 patients, with 3 patients showing a complete response, and 10 patients showing a partial response. Five patients had a stable disease for >6 months and two patients showed a progressive disease. No further therapies were prescribed in these patients. After a median follow-up of 36 months, the median duration of response was longer than 1 year.No patients reported grade 3 or higher treatment-related adverse events. Grade 2 adverse events were reported in five patients (25%). Adverse events included 12% pneumonia, 12% diarrhea, and an increase in alanine transaminase and aspartate transaminase. Grade 1 toxicity was recorded in 50% of the population .Overall, treatment was well tolerated, with 10% of patients discontinuing therapy not due to treatment-related adverse events (one hip fracture and one heart failure).At the time of the present analysis, twelve patients (60%) are currently on treatment with maintained disease control. The median duration of treatment was 15 months. The management of cSCCs requires a multidisciplinary approach. With the advent of Cemiplimab, the outcomes of patients with locally advanced/metastatic cSCC have significantly improved, and the indication for repeated local treatment  against systemic therapy should always be discussed in a multidisciplinary setting.Ultra-octogenarian patients with cSCC are potentially frail, and adequate selection for systemic treatment is mandatory. According to our experience, most patients with LAcSCC are elderly or frail with multiple comorbidities. Frailty is defined as an age-related condition of increased vulnerability to acute endogenous or exogenous stressors . TherefoBiological age does not represent a contraindication for Cemiplimab, but a comprehensive assessment of frailty is required in this population. So, scientific societies recommend CGA for patients older than 70 years old. However, more than 50% of all solid tumors and more than 80% of cSCC occur in patients older than 70 years old, making the systematic implementation of CGA difficult to perform in daily clinical practice . RecentlAmong screening tools, we used the G8 tool, which is a simple tool to routinely identify patients who should have a complete assessment in geriatric oncology .We found the G8 to be a very useful screening tool in the clinical practice to target a geriatric assessment, and it was more precise than the FRAIL score, which did not separate prognostic and comorbidity in our population. Indeed, most of our patients had 0 to 4 illness and weight loss as well as fatigue.Cemiplimab has significantly changed the clinical armamentarium of locally advanced/metastatic cSCC, as safety and objective response rates justify treatment also in elderly and frail patients, who were historically not candidates for cytotoxic chemotherapy .An adequate selection of patients before immunotherapy should be obtained in the context of an MDT. Some clinical predictors to define immunotherapy responses might be considered, such as the G8 score, obesity, polypharmacy, comorbidities, and a history of immunocompromised conditions .Concerning efficacy, our study confirms that the ORR in this special population of patients is similar to that reported in other real-world studies.As a retrospective case series, our data are in line with previous studies reporting data in the real-world.A previous Italian multicenter experience reported data on 131 patients with a median age of 79 years (range 19–95) and a history of immune suppression in nearly 20% of patients (9.2% with concurrent chronic lymphoproliferative disease and 8.5% with concomitant autoimmune disease) . In thisIn the German ADOReg Registry, 9 out of 39 patients were receiving immune-suppressive therapy during treatment with Cemiplimab. No significant differences between immune-competent and immunosuppressed patients  were registered, although these responses less often resulted in durable remissions . In thisAs in other solid tumors, complete response is associated with durable remission, likely sustained by significant changes in the tumor microenvironment . In the Due to the small number of patients, we cannot confirm a correlation between outcome and lactate dehydrogenase levels. According to other real-world experiences, the response rates among elderly patients with cSCC range from 32 to 80%. This might depend on patient characteristics, including age, disease stage, number of previous local treatments received, comorbidity, and immune- suppression. It must be stressed that in these published series, the range of age differs as well as the number of immunosuppressed patients and special population definition.In the Israelian experience of 102 patients treated with PD-1 inhibitors (Cemiplimab or Pembrolizumab), the response rate was higher than other real-life experiences (80%), with a complete response in 45.2% and partial response in 35.5% of the patients. According to this data, the authors supported immune checkpoint inhibitors to be amenable for use in elderly or frail patients with comorbidities .The phase IV CASE study is a big, non-interventional, survivorship and epidemiology study with Cemiplimab in a real-world setting which is currently ongoing. Patients’ enrolment is expected to be completed by September 2025 . PrelimiAlso, data from the French experience on 32 patients confirmed the other real-world experience  . In contA higher disease control rate (about 50%), in line with the previous experiences, was reported by Ríos-Viñuela . All theIn a retrospective study on 465 cSCC patients, both immunocompetent and immune-suppressed, Zavdy et al. demonstrated lower survival rates, higher rates of positive resection margins, higher recurrence rates, and multiple cSCC tumors in the immune-suppressed patients. Aetiologies for immunosuppression in this study included transplantation, chronic lymphatic leukemia, chronic kidney disease, psoriasis, rheumatoid arthritis, and systemic lupus erythematosus. Transplant recipients had multiple cSCC tumors (35%), with the highest number of primary tumors compared to controls, but also compared to all other immunosuppressed groups. The number of lesions and the advanced stages might be explained by the number of surgical procedures proposed .The strength of our series is that all patients were evaluated in an MDT setting specialized in skin cancer tumors, with long-standing experience, and the same systemic treatment was offered to all patients. The limitations of our study include its retrospective nature, the small sample size, and the length of follow-up. Further prospective data collection is warranted to collect more data on Cemiplimab treatment in this group of patients.cSCC is extremely common among ultra-octogenarian patients. Immunotherapy with anti PD-1 monoclonal antibody Cemiplimab can provide significant clinical benefits for patients with LAcSCC or metastatic cSCC who are not candidates for curative surgery or radiation. Elderly patients with comorbidities, or treated with concomitant immune suppression, are excluded from clinical trials. However, they represent a significant proportion of patients in everyday clinical practice. Indeed, not all centers can routinely perform a geriatric assessment.The G8 tool might help to identify elderly cancer patients who could benefit from CGA.In conclusion, our data suggest that Cemiplimab is also feasible in frail patients (both for age and/or comorbidities); a multidisciplinary approach is strongly recommended to correctly address patients to systemic treatment, if indicated.


Hematopoietic malignancies are the most frequent type of cancer in childhood. Recent advances in cancer treatment have significantly improved survival until adulthood. There is an extensive literature on the effects of cancer treatment on the gonadal axis in adult survivors of childhood cancer mainly focused on sperm production, but scarce information exists on the immediate impact of cancer and its treatment in boys.In this work, we determined the status of the hypothalamic-pituitary-testicular (HPT) axis function at diagnosis and the immediate impact of chemotherapy at the start of treatment in children and adolescents with hematopoietic malignancies.In a prospective study of 94 boys and adolescents with acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML) or non-Hodgkin lymphoma (NHL), we determined serum AMH, inhibin B and FSH to assess the gonadotrophin-Sertoli cell component of the HPT axis, and testosterone and LH to evaluate the gonadotrophin-Leydig cell component, at diagnosis and after 3 months of chemotherapy. Secondarily, the general health state was evaluated.In prepubertal boys, at diagnosis, AMH, inhibin B and FSH were lower compared to the reference population, reflecting an FSH-Sertoli cell axis dysfunction. After 3 months of chemotherapy, all hormone concentrations increased. At pubertal age, at diagnosis, AMH and inhibin B were lower compared to the reference population for Tanner stage, with inappropriately normal FSH, suggesting a primary Sertoli cell dysfunction with insufficient gonadotrophin compensation. The LH-Leydig cell axis was mildly disrupted. After 3 months of chemotherapy, inhibin B and AMH were unchanged while median FSH levels rose to values that exceeded the reference range, indicating a significant impairment of Sertoli cell function. Testosterone normalized concomitantly with an abnormal LH elevation reflecting a compensated Leydig cell impairment. General health biomarkers were impaired at diagnosis and improved after 3 months.The HPT axis function is impaired in boys with hematopoietic malignancies before the initiation of chemotherapy. There is a primary testicular dysfunction and a concomitant functional central hypogonadism that could be due to an impaired overall health. The HPT axis function improves during the initial 3 months of chemotherapy concomitantly with the general health state. However, in pubertal boys the dysfunction persists as shown by elevated gonadotropin levels after 3 months. Haematopoietic malignancies represent the commonest type of cancer in childhood : approxiWhile the assessment of the hypothalamic-pituitary-testicular (HPT) function is usually based upon the study of pituitary gonadotropins, testosterone secreted by Leydig cells and spermatogenesis in adults , in chilMost of the evidence of the effects of cancer treatment on the HPT axis relies on studies in adult survivors of childhood cancer and focuses mainly on sperm production ; scarce In this work, we aimed to determine the status of the HPT function at diagnosis in children and adolescents with haematopoietic malignancies and the immediate impact of chemotherapy at the start of treatment. In a prospective study of a large cohort of boys and adolescents, we longitudinally assessed serum levels of gonadotrophins and testicular hormones at diagnosis and after the induction phase of chemotherapy.We performed a prospective, analytical, study of a cohort of boys and adolescents diagnosed with haematopoietic malignancies at Ricardo Gutiérrez Children’s Hospitals, a tertiary paediatric public hospital in Buenos Aires, Argentina. Patients were recruited at diagnosis for long-term follow-up in the Unit of Haematology and referred to the Divisions of Endocrinology of Ricardo Gutiérrez, between March 2013 and February 2019. In this interim analysis, the status of the HPT axis was assessed at diagnosis and approximately after the first 3 months of chemotherapy, which corresponds to post-induction phase in the treatment of patients with ALL, acute myeloid leukemia (AML) and most non-Hodgkin lymphoma (NHL).rd and 4th editions of the World Health Organization Classification of Tumors of Hematopoietic and Lymphoid Tissues  . BrieflyAt diagnosis and after 3 months of chemotherapy, the patients were evaluated by a pediatric endocrinologist and were subjected to hormonal assessments. Blood samples were obtained during hospitalization between 07.00 and 10.00 AM, immediately after diagnosis. Both at diagnosis and after 3 months, most patients received oral feeding, and some received iv fluids to maintain a euvolemic state. Patients were grouped according to pubertal stages as defined by Marshall and Tanner . Testicu2, and expressed as standard deviation score (SDS) in boys > 2 years old. .The main outcome measures of the study were the serum concentrations of AMH, inhibin B and FSH to assess the gonadotrophin-Sertoli cell component of the HPT axis. Secondarily we evaluated serum testosterone and LH, to assess the gonadotrophin-Leydig cell component of the HPT axis, in pubertal patients. Serum hormone levels were expressed as absolute values and as standard deviation scores (SDS) based on age- and Tanner genital stage-matched reference ranges, from a sample of 421 apparently normal males using similar hormone assays, previously published by our group , 27 The All hormone assays were performed using fresh samples, except for inhibin B assay that was performed on frozen samples.®, Beckman-Coulter Co., Marseilles, France), as previously validated . Intraassay coefficients of variation were 14% for a serum inhibin B concentration of 111 pg/mL, and 9.8% for 479 pg/mL. Inter-assay coefficients of variation were 14% for a serum inhibin B concentration of 12 pg/mL, and 7.7% for 210 pg/mL. When serum inhibin B levels was undetectable, a value of 7.2 pg/mL, corresponding to the limit of quantification , was attributed.Serum inhibin B was determined using an enzyme-linked immunoassay specific for human inhibin B  as described . Intra- nd National Institute for Biological Standards and Control International Standard (NIBSC IS) 80/552 for LH and the 2nd World Health Organization International Reference Preparation (WHO IRP) 78/549 for FSH. Intra- and inter-assay coefficients of variation were 1.1% and 1.8%, respectively, for a mean LH concentration of 2.8 IU/L and 1.4% and 1.5% for a mean LH concentration of 16.9 IU/L. Intra- and inter-assay coefficients of variation were 1.0% and 4.2%, respectively, for a mean FSH concentration of 14.8 IU/L and 1.1% and 4.1% for a mean FSH concentration of 23.4 IU/L. When serum LH or FSH levels were undetectable, the value of the limit of quantification  was attributed.LH and FSH were determined using electro-chemiluminescent immunoassays  as described . The limSerum albumin and C-reactive protein (CRP) were measured using immunoturbidimetric assays in Cobas c501 (Roche), and hemoglobin was determined by a colorimetric method in Mindray BC-6800 Plus.P <0.05. All statistical analyses were performed using GraphPad Prism version 9.4.1 for Windows .Data distribution was assessed for normality using the Shapiro-Wilks test. Results are expressed as median and interquartile range. For comparisons of median SDS in patients at diagnosis with the theoretical median of 0 SDS in the reference population, we used the parametric one sample t test in case of normal distribution and, in the opposite case, the nonparametric Wilcoxon signed rank test was used. In the same way, depending on the distribution, paired t test or Wilcoxon matched-pairs signed rank test were used to compare between basal hormones and after 3 months of treatment in the individual follow-up of patients. The level of significance was calculated for groups ≥7 patients and set at The calculation of the study sample size was performed to determine the proportion of patients with Sertoli cell dysfunction up to 36 months of follow-up in the longitudinal cohort of boys and adolescents diagnosed with hematopoietic malignancies. All of the patients of the longitudinal study who had clinical and hormonal assessments at diagnosis and after 3 months of chemotherapy were included in the present study. Since this is an interim study of the whole cohort, a specific sample size calculation was not performed.Research was conducted in accordance with principles of the Declaration of Helsinki and local regulations on observational studies with human subjects. The study protocol was approved by the Institutional Review Board of the Ricardo Gutiérrez Children’s Hospital, Buenos Aires (# CEI 14.07). All patients or their parents, as appropriate, provided informed consent.Twenty out of 121 eligible patients were not included for the following reasons: 12 initiated chemotherapy treatment prior to the first hormonal evaluation, 7 had trisomy 21 and 1 had corpus callosum agenesis Figure 1Serum levels of AMH, inhibin B and FSH were measured to assess the functional status of the gonadotrophin-Sertoli cell component of the HPT axis. Given that reference hormone levels change during childhood and puberty, we analyzed our results using SDS. Nonetheless, absolute hormone levels are provided in When analyzed by diagnosis, baseline AMH was decreased in patients with SR/IR-ALL  but not in those with HR-ALL  or NHL   and AMH , whereas median FSH was within the reference range . Serum inhibin was below 0 SDS in 80.0% of the boys and 4 patients (26.7%) had levels below -2 SDS, while AMH was below 0 SDS in 83.3% but none was below -2 SDS. After 3 months of chemotherapy, median inhibin B  and AMH  remained below 0 SDS, with no significant changes compared to the basal state, whereas median FSH  increased significantly to abnormally high levels. FSH was above +2 SDS in 13 patients (54.2%)  with normal LH  at diagnosis. Serum testosterone was below 0 SDS in 83.3% of the patients and 3 patients (12.5%) showed serum testosterone below <2 SDS. After 3 months of chemotherapy, median testosterone increased significantly , while LH showed median levels above 0 SDS ; LH was above +2 SDS in 13 patients (54.2%)  and the seminiferous tubule (Sertoli cell) compartments, in agreement with previous findings in all cancer types  and in aIn conclusion, the HPT axis is impaired at diagnosis in boys with hematopoietic malignancies. There seems to be a combined hypogonadism, with a primary testicular dysfunction possibly due to gonadal infiltration by tumor cells and/or the effect of inflammatory factors on the gonad, and a concomitant functional central hypogonadism that could be due to an impaired overall health. The HPT axis function improves during the initial 3 months of chemotherapy, with certain nuances according to age and testicular compartment, concomitantly with the general health state.The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation.The studies involving human participants were reviewed and approved by Comité de Ética en Investigación, Hospital de Niños Ricardo Gutiérrez. All patients or their parents, as appropriate, provided informed consent.RG, RR, GD, LA, MGR and IB conceived the study design; JD, SP, LB, MB, MER, MEG, MS, LM, CF and PB collected clinical and laboratory data; JD, SP, LB, RR and RG analyzed the data; JD, RR and RG drafted the manuscript; All authors contributed to the article and approved the submitted version.


The mean number of devices per case was 4 ± 1.1 for the VP group and 2 ± 0.31 for the VPPF group (p < 0.001). No major complications were recorded in either group. The VPPF group had a significantly longer fluoroscopy time (42.8 min ± 14.2 vs. 25.4 min ± 7) and longer radiation dose ). Conclusions: Embolization for PCS resulted in pain relief in 90% of patients; the use of polidocanol did not demonstrate changes in the clinical outcome. The use of a VP alone was associated with decreased fluoroscopy time and radiation dose.Level of Evidence: Level 2. Purpose: To compare the safety and efficacy of vascular plug (VP) and vascular plug and polidocanol foam (VPPF) treatments for embolization in pelvic congestion syndrome (PCS). Materials and methods: A comparative, prospective, two-center study enrolled 50 women with PCS from January 2019 to January 2020. The patients were divided into two groups, and embolization was performed with VP (n = 25) and VPPF (n = 25) treatments. The mean age of the patients was 45.6 years ± 6.9. Three clinical parameters were assessed: abdominal pain, dyspareunia, and lower limb pain. The primary outcome , number of devices, procedure and fluoroscopy times, radiation doses, costs, and complications were compared. The participants were followed-up at 1, 3, 6, and 12 months. Results: At the 1-year follow-up, clinical success did not significantly differ between the two groups (VP vs. VPPF) regarding the improvement of the symptoms analyzed (pelvic pain, dyspareunia, lower extremity pain, and other symptoms ( Pelvic congestion syndrome (PCS), included in a pathology called pelvic venous disorders (PeVDs) in women , is a coPelvic venous congestion may be due to hormonal changes, venous valve insufficiency, or secondary to venous stenosis or obstruction ,6.The treatment of pelvic congestion syndrome (PCS) includes hormonal drug therapy and, in some cases, surgical ligation, including hysterectomy. However, endovascular embolization has been shown to be as effective as traditional surgery, while less invasive and better tolerated by patients ,8.Various embolizing agents have been used to embolize the gonadal or iliac veins involved in PSC. The purpose of this study is to compare a solid agent  (VP) with a solid agent (vascular plug) and a sclerosing agent (polidocanol 2% foam) (VPPF).This prospective comparative study was approved by the institutional review board  and was conducted in accordance with the principles of the Declaration of Helsinki. Written informed consent was obtained from all participants for both the procedure and the study. Eligible patients were assigned to two groups without randomization. The patients were consecutively assigned to one of the two groups without considering any differential factor to receive VP or VPPF treatment. All patients were recruited from January 2019 to January 2020. The primary outcome was clinical success subjectively assessed by patients. The secondary outcomes were complications with a need for re-embolization secondary to lack of improvement, recurrence of initial symptoms, or DVTUS revealing venous reflux or venous diameter > 6 mm.This study was conducted at two centers. The patients were referred by various specialists. All women underwent a Transvaginal Doppler Ultrasound (TVDUS) to diagnose pelvic varices and met the eligibility criteria . The diaThe sample size was determined using computer software  using prospective data from patients seen in 2019–2020 [A solid medium, such as the Amplatzer Vascular Plug (AVP) , and a liquid medium, a sclerosing agent, were used as embolizing agents. AVP II (Amplatzer) with diameters from 8 to 22 mm and lengths from 6 to 18 mm were used. Depending on its sheath, it requires a 5–8 F caliber catheter .2. The contents were mixed until a blend of homogeneous foam was achieved [Polidocanol  is a sclerosing and irritating liquid substance formed by the ethoxylation of dodecanol. Injected to treat varicose veins, it causes fibrosis within the varicose veins, occluding the lumen of the vessel and reducing the appearance of varicose veins. A mixture of 2% polidocanol with air and 1 cc of iodinated contrast Iopamiro 300  was used. The 2% polidocanol foam was prepared by connecting two 10 mL Luer lock syringes using a three-way stopcock containing 2% polidocanol  and 8 mL of COachieved .Two experienced interventional radiologists, both with >30 years of experience, performed all embolization procedures in both hospitals.The technique has been described by the authors in previous publications and has not changed substantially for plug embolization (Group VP). In all cases, the four venous axes were closed ,10.In the VPPF group, the existence of direct venous connections to the systemic venous circulation was ruled out. When these were found, the connection and flow were reduced by releasing a VP of the appropriate caliber. On a few occasions, it was required to close the connection with an occlusion balloon. Coaxially to the MPA 5F catheter, a 2.7 F 130 cm Progreat microcatheter  was introduced, and a foam of 2 mL of 2% polidocanol was mixed with 8 mL of air and 1 mL of iodinated contrast. This mixture was injected slowly until the vessel was obstructed, verifying that there was no migration of the foam towards the systemic venous circulation. Occlusion  was confirmed through venography performed after embolization . During Clinical success was defined as the relief of symptoms experienced before the procedure, including abdominal pain, dyspareunia, lower limb pain, dysmenorrhea, and urinary urgency . The subjective abdominal pain, dyspareunia, and lower limb pain was assessed with a VAS scored from 0 (no pain or symptoms) to 10 (worst pain or symptoms possible). Two categories were created: relief of symptoms (VAS score decreased by 4 points) and no improvement or worsening  [The patients in both groups had clinical follow-up at 1, 3, 6, and 12 months after the embolization procedure. The related symptoms were recorded, and subjective pain was assessed by the patients at each follow-up visit. When no improvement was observed, initial symptoms recurred, or DVT showed venous reflux or persistent venous diameter > 6 mm, re-embolization was scheduled.To assess quality of life, a telephone survey was conducted (at 12 months). The patients were asked about abdominal pain, sexual intercourse, and pain in the lower limbs. Each question was scored from 0 to 10 according to the following distribution: almost always = 0, often = 1–3, sometimes = 4–6, occasionally 7–9, and never = 10. At the end of this study, a survey was conducted, including regarding the quality of the telephone service, with patients scoring their satisfaction with the procedure: the treatment received and the possibility of recommending it to other patients from 0 to 10.p < 0.05 was regarded as significant in all two-sided tests of significance. Using IBM SPSS Version 21.0 , data were processed and examined.By applying the Kolmogorov–Smirnoff test, normality was evaluated. The mean, standard deviation, and range were used to express quantitative variables. Total events and percentages were used to convey qualitative data. The parametric Student t test and the nonparametric Mann–Whitney U test were used to assess continuous data. For categorical variables, the Thec2 test and Fisher exact test were employed. A scatterplot was used to represent time data, and a line chart was used to show how the VAS score changed over time . Fifty patients were randomly assigned to the two groups. Except for those excluded in the first month, no patients were lost at 3, 6, or 12 months of follow-up, and all 50 patients were analyzed. See p < 0.0001). The total procedure time, total fluoroscopy time, and radiation dose were significantly lower in the VP group compared to the VPPF group . Tin toto achieve total occlusion of the four veins in the VP group, a mean of 4.2 plugs was used. The sclerosing substance was never used in this group. In the VPPF group, a mean of 2.2 plugs were used, and all four venous axes were sclerosed in 21 (84%) patients. Sclerosis in the hypogastric veins was ruled out in four patients because they had a very wide communication with the external iliac system  presented pelvic pain with a mean intensity VAS of 6.4 ± 3.1 (range 0–10) with a duration of 40 min ±27 h (range 0–72 h) that required treatment with Metamizole and/or Tramadol. Higher VAS values and longer duration of pain were observed in the VPPF group A,B.Clinical success was achieved at 1-year follow-up in all 50 patients. At 1-year follow-up, clinical success did not differ significantly between the two groups (VP vs. VPPF) in terms of improvement in the symptoms analyzed .In both groups, the relief of symptoms was found in the VAS for abdominal pain and dyspareunia, pain in the lower limbs, and other symptoms (there was a decrease of >4 points with respect to the pre-treatment score) in 88%, 96%, and 48% of patients, respectively. The VAS for lower limb pain and other symptoms showed no worsening but no change in 52% of patients.Two patients’ symptoms in each group did not improve (score <3 points) in a corresponding way when the four values  were analyzed compared to when only abdominal pain was analyzed. Of the patients whose symptoms did not improve, two of them, one in each group, underwent a second embolization at 5 and 10 months, respectively; despite this, the symptoms did not improve substantially.The satisfaction score, personal treatment, and recommendation interest at the end of this study, according to the quality questionnaire carried out by telephone, can be seen in It has been described that embolization produces total or partial improvement in symptoms in between 60 and 100% of patients diagnosed with pelvic venous disorders (PeVDs) . Despitep > 0001) and therefore a higher air kerma and DAP . Another limitation is that clinical success, referring to abdominal pain and pain and discomfort in the lower limbs, is difficult to measure objectively. A subjective scale, such as the VAS, can be influenced by other concomitant disorders and environmental or psychological conditions .While complications arising from venous embolization are infrequent, certain complications can bear clinical significance. Both minor and severe complications are plausible and may manifest during the surgical procedure, shortly thereafter, or during the postoperative follow-up period. This can include vein thrombosis, entry-point hemorrhage, changes in heart rate due to right heart passage, inadvertent release of embolizing agents into unintended veins, material migration, venous rupture, and iodinated contrast adverse reactions.Of the studies encompassed within the systematic review by Daniels et al. , six stuIn a survey conducted by Leal Monedero  in 2006,Given the paucity of research pertaining to its investigation, the precise impact of vein embolization on fertility remains unknown, although instances of pregnancies after embolization are well documented ,28,29.The main long-term complications include migration to the lung, thrombosis of the access vein, foreign body reactions, and compression-induced neuralgia. Conversely, immediate complications primarily entail pain, abdominal discomfort, and the unintended migration of embolizing agents.Henceforth, whether employing venous plugging (VP) or venous plugging plus polidocanol foam (VPPF), the practice of venous embolization as a therapeutic intervention for pelvic congestion syndrome (PCS) demonstrates equivalent levels of safety and efficacy. Both modalities yield a substantial alleviation of pelvic symptoms associated with PCS. Clinically, the outcomes of both procedures reveal a comparable attainment of varicose vein occlusion.In a direct comparison of the VPPF and VP techniques, it is evident that the VPPF procedure requires a longer duration and exposure to radiation. Furthermore, the occurrence of postprocedural abdominal discomfort is notably more frequent in cases of VPPF as opposed to VP, consequently requiring analgesic intervention as a usual measure.


The number of patients reaching a ≥20% output reduction was 10/19 (52.6%) treated vs. 3/20 (15.0%) untreated patients (p = 0.013) at 1 month and 12/19 (63.2%) vs. 6/20 (30.0%) (p = 0.038) at 6 months, respectively. Participants with a clinically relevant output reduction at 6 months had a significantly lower baseline weight and BMI. Energy parenteral supply significantly decreased, while infused volumes, oral energy, and fluid intakes slightly decreased, though not significantly. This pilot study supports liraglutide benefits in ostomy/fecal output early after surgical gut resection in SBS patients, particularly in those with lower baseline weight values.Liraglutide, a glucagon-like peptide-1 agonist, has been shown to have beneficial effects on fecal output in short bowel syndrome (SBS) by small human studies. Its potential effects early after gut resection are not known. In this pilot observational study, we described the 1- and 6-month liraglutide effects in 19 adult patients with a new SBS diagnosis within 1 month after surgical resection. Stomal/fecal and urinary outcomes, serum/urinary electrolytes, and body composition were assessed. Both within-group differences and between-group comparisons with 20 SBS patients refusing liraglutide treatment were evaluated. The main liraglutide-related side effect was mild nausea, except in one patient, who experienced severe nausea/vomiting. The median ostomy/fecal output was significantly reduced by −550 mL/day after 6 months of treatment ( Chronic intestinal failure (CIF) is the long-lasting reduction of gut function below the minimum necessary for nutrients and/or water/electrolytes absorption, requiring intravenous supplementation to maintain body health and function, and nutritional homeostasis .Short bowel syndrome (SBS) is the most common cause underlying CIF and derives from extensive surgical resection due to various diseases, such as ischemic, neoplastic, inflammatory, or congenital bowel diseases . PatientFollowing extensive gut resection, depending on the extent and location of the resection, the absorptive capacity and length of the remnant bowel, the underlying disease, and the dysfunction in the remaining intestine, patients experience malabsorption of fluid, energy, and nutrients . Based oContrary to the large number of studies on GLP-2, very few and small human studies have assessed the effects of GLP-1 agonists on ostomy output in SBS patients ,11,12. OIn the present open-label pilot observational study, we aimed to describe the 1- and 6-month effects of liraglutide on many clinical outcomes in SBS patients early after surgical intervention. Comparisons with an untreated group of SBS patients were performed too.This was a single-center pilot observational study. All the adult patients with a new diagnosis of SBS after surgical gut resection requiring parenteral nutrition (PN) and followed up at the Intestinal Failure Unit of the “Città della Salute e della Scienza” Hospital of Torino, a tertiary referral center for CIF support, from 1 January 2017 to 30 June 2022 were evaluated for suitability for liraglutide therapy.The inclusion criteria were age ≥18 years, surgical gut resection within 1 month, new diagnosis of SBS confirmed by radiological examination, need for PN, and ongoing treatment with antisecretory agents (pantoprazole or lansoprazole) and antidiarrheal medication (loperamide) for at least 3 weeks at the maximum dosage tolerated by the patients. Exclusion criteria were active neoplastic disease and/or being under antineoplastic treatment within the previous 5 years, inability to give informed consent, critically ill patients or patients with a <6-month life expectancy, presence of renal, liver, or cardiac diseases, or contraindications to GLP-1 agonists . These criteria were formulated to include a relatively homogeneous group of patients and to exclude conditions potentially interfering with fecal output, fluid and electrolyte balance, life expectancy, and the possibility to perform an adequate follow up of the patients. Out of 59 patients with a new diagnosis of SBS, 46 satisfied the above-reported criteria. Of them, 22 accepted liraglutide treatment, and 24 refused the injections.Information on untreated patients was obtained from the database of the Intestinal Failure Unit to compare ostomy output and fecal losses to those of liraglutide-treated participants. Complete data were available for 20 out of 24 untreated patients and were collected at comparable times to those of the liraglutide-treated patients, i.e., within 1 month after surgical resection (enrolment) and 1 and 6 months after enrolment .All patients gave their informed written consent to the collection and processing of their data. In addition, patients signed a form accepting the off-label administration of liraglutide. This module explained in detail the potential risks and adverse events related to the administration of the drug. The study was approved by the local ethics committee (protocol number 595/2022); all the procedures were in line with the principles of the Declaration of Helsinki.The primary outcome was the assessment of the ostomy output and fecal losses of the patients at 1 and 6 months after liraglutide treatment. The secondary outcomes were changes in body weight and serum and urinary electrolytes, number of nights/week of PN, and volume of the infused solutions. Finally, fecal outputs of liraglutide-treated patients were compared with those of the untreated group.The Intestinal Failure Unit database contains information about all patients with CIF since 1985 . PatientDemographic, clinical, and laboratory data were extracted from the database.All patients received nutritional support according to their individualized needs. Personalized daily fluid and nutritional parenteral programs were balanced according to nutritional requirements, considering body weight, 24 h urine and ostomy output, oral caloric intakes, and laboratory exams in line with a standardized procedure by the same two trained physicians.PN was administered by an intermittent schedule at night for 10–16 h/day. All patients were given a diet restricted in fiber and lactose with an energy amount appropriate for the individual needs and corrected for the estimated fecal energy losses. The intake of both oral hypotonic and hypertonic fluids was restricted. Nutritional care, centralized laboratory and radiological exams, and handling of central venous catheters and of PN complications were performed in line with recommendations [Specialized nurses visited patients at their home monthly (or as needed) to ensure adequate home care support. Patients were contacted by telephone every 15 days by the physicians of the Intestinal Failure Unit, while hospital follow-up visits were performed every 30 days.Body weight and height were measured, with the patients wearing light clothes and no shoes, by a mechanical column scale (SECA model 711) and a stadiometer  after fasting in the morning, after urination, and after emptying stoma bags or defecation. Body composition was determined by single-frequency bioelectrical impedance using the manufacturer’s equations . The phase angle (measured in degrees) was obtained by bioelectrical analysis and is an indicator reflecting cell physiological functional level and volume. Arterial blood pressure values were measured on the left arm, while sitting, after at least 10 min of rest, with a mercury sphygmomanometer with appropriate cuff sizes . Blood and urinary samples were collected after an overnight fast. The laboratory methods were centrally measured according to standardized methods. Energy intakes were assessed during each visit by the same trained dietician by asking the frequency and portions of all the foods consumed in the last 7 days before the visit.®; Novo Nordisk, Bagsværd, Denmark; ready-to-use 6 mg/mL prefilled pens) was subcutaneously self-injected by the patients in the abdomen or thigh every morning. Starting dose was 0.6 mg/day for 1 week, followed by 1.2 mg/day for another week and then 1.8 mg/day until the end of the observation period. The dose schedule could be modified based on the individual tolerance to consist of a slower dose escalation (every 2 weeks) or maintaining the maximum tolerated dose. During the follow up, the dose of liraglutide could be changed according to the clinical response. Compliance to liraglutide treatment was assessed during each telephone call and visit.Liraglutide  occurred .t-test or Wilcoxon matched-pairs test, as appropriate. Between-group differences were analyzed by Student’s t-test or the Mann–Whitney test. A multiple logistic regression was employed to evaluate the association between liraglutide response and clinical variables.Results are reported as mean ± SD or percentages. Within-pair differences were tested by one-way repeated measures ANOVA or, in the case of non-normally distributed values, by Friedman ANOVA. Variable values at 1 month and 6 months were compared to baseline values by paired Out of the 22 enrolled participants, one patient experienced severe vomiting and nausea, leading to the early discontinuation of liraglutide; two other patients suspended the drug between the third and sixth months due to unwillingness to continue subcutaneous injections. The data of 19 patients were therefore analyzed.Most participants were middle-aged males with end-jejunostomy . The undAll patients started PN after the surgical resection and were on the maximum tolerated doses of antisecretory and antidiarrheal medication. These dosages were not changed during the 6-month period of observation.No patient reported either hypoglycemia or serious adverse effects. During follow up, liraglutide dosage could be changed in order to achieve the best balance between maximizing therapeutical effects and minimizing side effects. The main side effect was mild nausea  reported by eight patients and requiring the reduction of the drug dosage to 1.2 mg. After this therapeutic change, the symptom resolved.During the follow up, a slight, not significant increase in body weight, BMI, and fat-free mass, cellular mass, phase angle, and urinary 24 h sodium excretion was observed . The urip = 0.016) and baseline BMI . In a logistic regression model, the inverse association between response to liraglutide and baseline body weight  remained statistically significant after adjusting for gender but not after adjustments for other variables. Response to liraglutide and baseline BMI were inversely associated after adjustments for age .Ten and twelve patients showed a ≥20% reduction in ostomy/fecal output at 1 month and 6 months, respectively. Both at 1 month and at 6 months, the presence of a clinically relevant change in the ostomy/fecal losses was associated with neither age, gender, small bowel or colon length, anatomic classification, nor with the presence of the ileocecal valve. Participants with a clinically relevant reduction at 6 months had a significantly lower baseline body weight . The median ostomy/fecal output of these patients was 2200 mL/day at enrolment, 2000 mL/day at 1 month, and 2000 mL/day at 6 months (p for trend = 0.09) (p = 0.04) and −150 mL/day . The number of patients reaching a ≥20% reduction in ostomy/fecal output was three  at 1 month and six  at 6 months, respectively. No association between the ≥20% output reduction at 6 months and baseline weight or BMI was found in these patients. At 6 months, six patients were weaned to ≥1 nights/week PN , but three patients required more nights/week of PN , and none was fully weaned (p = 0.64 with respect to liraglutide-treated patients).The clinical characteristics of the 20 untreated patients are reported in  = 0.09) . The medy weaned  (p = 0.6Liraglutide treatment early after surgery was associated with a reduction in the ostomy output or fecal losses in patients with SBS.GLP-1 is quickly released after a meal from the distal bowel and enhances insulin production and secretion, decreases glucagon secretion, and increases glucose uptake and glycogen synthesis in peripheral tissues, thus reducing serum glucose levels ; furtherAccording to the literature , the effWe cannot exclude that the observed reduction in the ostomy/fecal losses may have been a consequence of an early innate intestinal adaptation, spontaneously occurring within the first 2–5 years after gut resection, due to the hypersecretion of the pro-adaptive distal hormones, gradually leading to a restoration of the intestinal absorption . This prThis was a small, open-label observational study, and, by its nature, it had many limitations. First, the number of studied patients was small; however, previous studies had a much smaller sample size. Furthermore, differently from others, we first assessed the effects of liraglutide administered early after surgery. In other studies, liraglutide was given to patients continuously dependent on PN for at least 6 months or longer ,9. We deThe dosing schedule of liraglutide was not defined a priori, but was conditioned by the patient’s clinical response since this was a real-life study, and patients were treated according to standards of care. Nevertheless, our study took place at a tertiary referral center for CIF support, with skilled professionals and proper organizational and logistic facilities. The adjustments in PN were guided by a standardized approach, according to guidelines, based on body weight, 24 h urine and ostomy/fecal output, oral caloric intakes, and laboratory exams. Unfortunately, in our hospital, variables and exams of potential interest, such as fecal energy and macronutrients, hormone profile, tests for gastric emptying, and analysis of the histological changes in the gut mucosa, are not routinely performed. Finally, a mix of residual anatomy, including patients with at least a part of a colon in continuity, may have affected the fecal volume losses of our patients. A randomized controlled trial on a large number of participants would allow an adequate evaluation of the effects of liraglutide by overcoming the potential confounding effects of an observational study without a control group.In conclusion, this small pilot observational “real-life” study supported the benefits of liraglutide in terms of ostomy/fecal output early after surgical resection in SBS patients. Unexpectedly, the greatest benefits were observed in patients with lower baseline weight values. These results are worth testing by a randomized, placebo-controlled trial with a larger sample size.


Neurobrucellosis (NB) presents a challenge for rapid and specific diagnosis. Next-generation sequencing (NGS) of cerebrospinal fluid (CSF) has showed power in detection of causative pathogens, even some infrequent and unexpected pathogens. In this study, we presented 8 cases of NB diagnosed by the NGS of CSF.Between August 1, 2018 and September 30, 2020, NGS was used to detect causative pathogens in clinically suspected central nervous system (CNS) infections. Data on demographics, clinical features, and laboratory tests, imaging results and NGS results were collected and reviewed.Brucella was rapidly detected using NGS of CSF within 1–4 days, despite those eight patients had variable medical history, disease course, clinical manifestations, laboratory tests and imaging findings. NGS showed the sequence reads corresponded to Brucella species were 8 to 448, with genomic coverage of 0.02 to 0.87%. The relative abundance was 0.13% to 82.40% and sequencing depth was 1.06 to 1.24. Consequently, patients were administered with 3 to 6 months of doxycycline, ceftriaxone and rifampicin, double or triple combination, supplemented with symptomatic therapy and were fully recovered except for case 1.Among the presented 8 patients, Brucella in a prompt and specific manner, and can be considered for first-line diagnostic use in practice.NGS of CSF provides a powerful tool in detection of  Brucellosis is the most prevalent zoonotic infection, with a population-based incidence of about 21.5/100,000 person-years . NeverthThe main problem with NB is the inadequate- or under-diagnosis, and the primary causes is the non-specific clinical manifestations that NB can presents as encephalitis, meningoencephalitis, polyneuropathy, subarachnoid hemorrhage, radiculitis, myelitis, and psychiatric manifestations et al. , 5. The Next-generation sequencing (NGS) is an emerging sequence-based pathogenic microorganism identification technique, and is increasingly used to diagnose the infectious disease of the central nervous system (CNS) –6. RevieGiven above, in this study, we presented 8 NB cases quickly diagnosed by the NGS of the cerebrospinal fluid (CSF) in the setting of a tertiary referral center in China, reported as follows.In this study, eight consecutive patients clinically suspected of having NB were admitted to the Department of Neurology of the second Hospital of Hebei Medical University, a tertiary referral center in Hebei province, China, between August 1, 2018 and September 30, 2020. As a part of a research project aimed at using NGS of CSF to detect pathogens in patients with clinical suspected CNS infectious diseases, this study of rarely seen NB cases gained particular attention, and our team prospectively and carefully collected the relevant data during the whole diagnosis and treatment process. The recorded data included demographic characteristics, clinical, radiological and pathogenic findings, and the treatment and outcomes. The protocol of the research project was approved by the Ethics Committee of the Second Hospital of Hebei Medical University (2020-R527), and in accordance with the principles laid down in the Declaration of Helsinki. Before the commencement of this study, each patient involved provided his/her written informed consent to participate.Patients with following clinical presentations, epidemiology and laboratory test findings were suspected of having NB: 1, history of fever; 2, clinical presentations as altered mental status , epileptic seizure and neck rigidity; 3, CSF findings of leukocytosis, decreased glucose level and rising protein level; 4, epidemiology: recent close contact with livestock, animal products or inhabitants in the affected area before the onset of the disease.During the lumbar puncture, an additional 2 ml of CSF was taken from each patient and placed into the sterile test tube (SARSTEDT Germany), stored in the refrigerator at − 80 °C within 30 min.The 2 ml CSF samples were then transmitted on dry ice for PACEseq NGS detection . QIAamp DNA Micro kit  was applied to extract DNA following the manufacturer’s manuals. Then, DNA libraries were constructed using the QIAseq™ Ultralow Input Library Kit for Illumina . All constructed libraries were assessed for quality using Agilent 2100 Bioanalyzer  and Qubit fluorometer 2.0 . Finally, the qualified DNA libraries were sequenced on an Illumina NextSeq 550 system platform .ftp://ftp.ncbi.nlm.nih.gov/genomes/) using Burrows-Wheeler Aligner software. The reads number and reads per million (RPM) of each detected pathogen was calculated. For detected microbes, including bacteria (mycobacteria excluded), fungi (Cryptococcus excluded), and parasites, a positive NGS result was given when its coverage ranked top10 of the same kind of microbes and absent in the negative control  or when its ratio of RPM between sample and NTC (RPMsample/RPMNTC) > 10 if RPMNTC ≠ 0. For viruses, M. tuberculosis, and Cryptococcus, a positive mNGS result was considered when at least 1 unique read was mapped to species level and absent in NTC or RPMsample/RPMNTC > 5 when RPMNTC ≠ 0.After the sequencing data is offline, adapter, low-complexity, low-quality and duplicated reads and short reads (length < 35 bp) were removed from the raw data of each library to generate high-quality data, using an in-house program. The human host DNA reads were also filtered out by alignment to human reference database (hg38). The clean reads were finally aligned to the Microbial Genome Databases  were in summer, case 6 and 8 in spring, case 1 in autumn and case 2 in winter. Five cases  had acute onset, and three  had chronic course. All had none history of comorbidities except case 1 who had tuberculosis. Four patients  had a history of contact with cattle and sheep . The general characteristics and clinical manifestations varied greatly between them and were summarized in Table 12O). Routine CSF examination showed elevated white blood cell (WBC) count (68 to 442 × 109/L), with mononuclear cells in absolute predominance (75–90%), elevated protein level (0.8–4.7 g/L) in all except for case 8; reduced glucose in case 1, 2, 4 and 7 (0.91 to 2.40 mmol/L). CSF cytology analysis was performed in six cases, with lymphocytic predominance in case 3, 4, 5 and 7 and no cytological abnormalities in case 8. CSF culture showed negative results for all the 8 cases.HIV antibody, syphilis antibody and TORCH tests showed negative, also the cerebrospinal fluid (CSF) assays for acid-fast staining, smear and culture, among these 8 patients. Five patients  had increased cranial pressure  and standard agglutination test (SAT) were positive in case 1, 3 and 7, RBPT positive in case 4, and SAT positive in case 6. Only in case 8, the blood culture identified the Brucella infection , and after 1 to 4 days the reports returned. A total of 20 species of microorganisms in 13 genera were detected. The proportion of human host nucleic acid sequences was 73.85–99.95% . After excluding background sequences, the sequence reads corresponding to 0 days, aAfter definite diagnosis, all the eight patients were all administered with 3 to 6 months of doxycycline, ceftriaxone and rifampicin, double or triple combination, supplemented with symptomatic therapy. Seven patients had symptoms significantly relieved, and the reexamination of CSF showed substantially decreased WBC and protein level, and were discharged to continue treatment. The condition of the case 1 was not improved, and he gave up continuing treatment and was discharged voluntarily.In this study, we presented 8 cases of NB in patients who had clinically suspected symptoms or signs, which were definitely and promptly diagnosed with NB using NGS of the CSF. NGS exhibited its superiority over the traditional methods in diagnosis of NB, allowing the early, prompt, high-sensitive and high-specific diagnosis to be realistic. As far as we know, this is the largest case series where NB was diagnosed by NGS of the CSF.Occupational exposure to animals plays the main factor in the more exposure of males to the incidence of brucellosis , which cBrucella) or inaccuracy [Currently, diagnosis of NB is primarily dependent on serology, microbial culture, PCR and first-generation sequencing, but various problems exist relating to the lag , but that did not affect the power of NGS in detection of Brucella, suggesting its robustness regardless of the course of disease.Compared with traditional molecular detection methods, meta-genomic NGS has the advantages of high throughput, fast and accurate diagnosis for pathogens, which not only overcomes the limitation of targeted diagnosis by primer design in PCR, but also can obtain drug resistance mutation information and virulence genes to assist diagnosis . All thenfection . Additionfection –20. In tBrucella infection in central nervous system.There were several case studies that examine the NGS of CSF to diagnose the NB, also showing consistent excellent performance. Fan et al.  retrospeAs an emerging technique, NGS faces with a very common problem, contamination, as described in ours and previous studies. This problem is intractable because the sources for contamination are various, such as skin or body flora during lumbar puncture, cross-contamination between samples during preparation of NGS library , or the There were several limitations to this study. First, the limited number of consecutive patients might affect the results; however, it is unrealistic to conduct a large sample study before NGS technique is extensively used in practice. Even so, the consistent promising findings in this study show the great and particular potential in clinically diagnosis of some infrequent infectious cases of CNS. Second, the single-center design might compromise the generalizability of the results, especially in case of different background microorganisms in individual institutions and contamination. So, the test findings should be comprehensively analyzed and treated, combined with the clinical presentations, laboratory findings and imaging characteristics.In conclusion, meta-genomic NGS is a promising technique in prompt and precise diagnosis of infectious disease of the CNS, like NB, through our 8 diagnosed consecutive cases using NGS of CSF. This technique aids to guide the targeted therapy and improves the management of NB, leading to an improved prognosis. Despite that, lack of standardized protocol and the contamination from various resources may confound the diagnosis, and the results should be treated comprehensively.


Meanwhile, a bioinformatic analysis of PHY gene family members was performed, and the results showed that the five StPHYs were distributed on chromosomes 1, 2, 5, 7 and 10, with amino acid counts ranging from 704 to 1130. StPHYs genes have abundant light-responsive elements. Also, the expression patterns of StPHYs were dramatically induced by red light. Additionally, a correlation analysis showed that the GAs accumulation was significantly correlated with StPHYs expression. This research is useful for comprehending the metabolism of GAs regulated by light and monitoring food safety in potatoes.Potatoes are the fourth major food crop in the world. Higher levels of glycoside alkaloids (GAs) lead to detrimental effects on the edibility and processing qualities. GAs are largely influenced by light; however, the mechanisms of this regulation by light are not well understood. By analyzing the bioinformatics of the phytochrome genes (PHYs) in potatoes, its expression level, the content of GAs and the correlation between them under different lights, this study aims to reveal the specific mechanism of light-regulated GAs accumulation and provide a theoretical basis for improved potato processing. Results based on high-performance liquid chromatography and imaging mass microscopy showed that red light induced a significant increase in  Solanum tuberosum L.) is an annual dicotyledonous herb in the Solanaceae family that is native to South America. As the world’s fourth largest food crop, it is planted all over the world. It is rich in carbohydrates, essential amino acids, vitamins and minerals [Solanum nigrum L. In the early 19th century, the isolation and identification of more than 100 GAs from plants were performed, and currently about 46 GAs, such as α-chaconine, α-solanine and α-tomatine, have been found in solanaceous species, for example, potatoes and tomatoes [The potato , cryptochrome (CRY), phototropin (PHOT) and ultraviolet-B receptor (UVR) . The phytostable . Upon actostable .Arabidopsis [Arabidopsis, both phytochrome genes have been found to contribute to chloroplast formation in R and FR light [PHY A has been reported to participate in regulating the carbon flux through a series of metabolic pathways, like glycolysis and the tricarboxylic acid (TCA) cycle . Also, Kbidopsis . In ArabFR light . Under lFR light . AlthougPotato cv. Atlantic, Favorita, EV and Gan Yin No. 3 were grown in the Potato Research Institute in Dingxi city, Gansu Province, China. These cultivars were cultivated in an experimental fileds that are uniformly managed. Organ samples and tubers were collected and harvested in August and September in 2023, respectively. After that, the obtained samples were taken to the laboratory and stored at 4 °C for experimental analysis.One hundred and fifty potato tubers were washed and air dried at room temperature (22 ± 2 °C). Then, these tubers were divided into three groups randomly. For red-light treatment, the potato tubers were placed in a light-tight box and provided with constant light using light-emitting diodes (LEDs) for up to 12 days. The wavelength of red light was 660 nm with ± 10 nm bandwidth; the other group was exposed to white light (400–680 nm), and tubers placed in darkness were used as control. Samples from the tuber epidermis of about 3–4 mm depth and the organs of roots, stems, leaves and flowers were immediately frozen in liquid nitrogen and then stored at −80 °C for glycosidic alkaloid extraction and total RNA extraction. The samples with three replications were taken at 0 d, 3 d, 6 d, 9 d and 12 d, and stored for further use.v/v, 1:10) ammonia. Then the precipitate was dried under vacuum conditions, dissolved in a tetrahydrofuran/acetonitrile/20 mmol KH2PO4 solution with a ratio of 1:3 and filtered through a microporous membrane for HPLC analysis. HPLC separations were performed with a Shimadzu HPLC system coupled with a chromatographic column of Inertsil NH2  using a mobile phase consisting of acetonitrile/KH2PO4  at a flow rate of 1 mL/min. Standards of α-chaconine of 95%  and α-solanine of 95%  were used for the LC analyses.Glycoside alkaloids were extracted and detected according to the method of Zeng et al. 2017) with some modifications [ with som® Version 1.30 software .The imaging MS analysis of GA accumulation was based on the method of Deng et al. (2021) with some modification . EpidermStPHY genes was carried out as described previously by Zhang et al. (2020) [StPHY genes, and then bioinformatically analyzed them through the online website according to the amino acid sequences of PHYs. MEGA X was used to construct the phylogenetic tree; the TBtools v1.09832 tool was used to draw the chromosome distribution map of the gene and analyze the StPHY genes structure, the promoter cis-acting elements of StPHYs and the promoter cis-acting elements.Identification of the . (2020) . Based oStPHYs were analyzed using a 2 × SYBR Green qPCR Mix kit . RNA was extracted from the roots, stems, leaves, flowers and tubers of different potato varieties , and then the RNA was reverse-transcribed into single-stranded cDNA using a reverse transcription kit . Primers for the PHY family genes were designed using Primer Premier 5 software, and the sequences of the primers are shown in detail in StPHYs were calculated using the 2−ΔΔCt method [The relative expression of the t method .p < 0.05. A correlation analysis was carried out in Origin 2021 software.All the data are presented as the means ± SE, which were also analyzed for statistical significance by Ducan’s multiple range test using SPSS 20.0 software at α-chaconine, dehydrosolanine and α-solanine under different light treatments can be clearly viewed in the optical images of potato periderm that are shown in We observed variations in the spatial distribution and relative content changes of four glycoside alkaloids in the potato periderm. The distributions of four kinds of GAs, including dehydrochaconine, The regulation of GA content by light is particularly obvious in contrast with other environmental factors, and photoreceptors can accurately sense light signals in the environment and regulate their synthesis. Phytochrome, as the red photoreceptor, plays a major role in potato photomorphogenesis . TherefoStPHYA, StPHYB, StPHYB2, StPHYC and StPHYE, whose relevant physicochemical properties are shown in detail in StPHYs gene family members encoded proteins with amino acid numbers ranging from 704 to 1130 and molecular weights ranging from 77,442.38 to 125,782.06 Da, among which the StPHYE protein had the lowest amino acid number and the smallest molecular weight. The CDS lengths of StPHYs genome members range from 1467 to 2040 bp, and the isoelectric point (PI) sizes range from 5.68 to 5.8, indicating that StPHY proteins are acidic proteins. The total average hydrophobicity index ranges from −1.071 to −0.111, indicating a low hydrophilicity. Subcellular localization showed that the StPHYs genes are all nuclear genes (A search of the potato genome-wide database (Solanum tuberosum v6.1) for PHY genes yielded five members of the PHY gene family, namely ar genes .StPHYs family members, all of which are break genes and separated by introns. The StPHYs family possesses six conserved domains: PAS_2 (PF08446), PAS (PF00989), HATPase_c (PF02518), GAF (PF01590), PHY (PF00360) and HisKA (PF00512). All members contain the typical PAS-GAF-PHY domain at the N-terminus and three structural domains: PAS_2, HisKA and HATPase_c  search with other PHY species. A total of 36 PHY proteins were obtained from 10 species, including Solanum tuberosum, Arabidopsis thaliana, Pisum sativum, Nicotiana tabacum, Sorghum bicolor, Zea mays, Oryza sativa, Solanum lycopersicum, Vitis vinifera and Capsicum annuum. The phylogenetic evolutionary tree was constructed and is shown in StPHY proteins were closely related to those of Solanaceae. StPHYB is more closely related to SlPHYB1, SlPHYB2 and NtPHYB, whereas StPHYA is closer to SlPHYA and CaPHYA; StPHY is closer to SlPHYE; and StPHYC is closer to SlPHYC and VvPHYC  were predicted in Tbtools v.1.09876 software. A total of 16 elements were screened and identified, and could be categorized as light-responsive cis-elements, phytohormone-responsive cis-elements, environmental-stress-responsive cis-elements and transcription factor binding sites. Hormone response elements include the auxin element, the salicylic acid response element, the gibberellin response element and the abscisic acid response element. Light-responsive cis-elements were most abundant in the environmental stress response. The results revealed that light-responsive elements had the largest proportion in the five potato StPHY family genes. The ABA-responsive cis-elements were the most numerous among the phytohormone-like responses ; the correlation coefficients were 0.74 and 0.75, respectively. In addition, StPHYC was also significantly correlated with α-chaconine and α-solanine contents, with a correlation coefficient of 0.68 and 0.7. However, StPHYA and StPHYB2 expressions were negatively correlated with GA content (p < 0.05). It is hypothesized that the red-light receptor protein StPHYs is involved in GA accumulation in potato tubers and it may be a major regulator (The correlation between PHY gene expression and egulator .2 s−1, the accumulation of GAs increased with increasing light intensity, whereas if it was more than 750 μ mol∙m2 s−1, it decreased with increasing light intensity [α-chaconine is a prevalent GA, as described by Friedman [Light is an important environmental factor in the regulation of potato growth and the content of GAs . The durntensity . Interesntensity . The resFriedman . In the Friedman .Arabidopsis belong to the PHYA, PHYB, PHYC, PHYD and PHYE subfamilies, respectively, with the PHYB and PHYD subfamily genes showing a higher degree of similarity, which is explained by the fact that the PHYD subfamily was derived from the PHYB subfamily in the last whole-genome duplication event in Arabidopsis [Oryza sativa and Zea mays, while there are five phytochrome in potatoes, including PHYA, PHYB, PHYB2, PHYC and PHYE, which are localized on five different chromosomes (StPHYs. With the exception of the StPHYC gene, all of the genes had high gene densities at both ends of the chromosome. All the phytochromes share a photosensory core module, consisting of PAS, GAF and PHY domains and a bilin chromophore [Phytochromes are important in light-induced development throughout plant growth, including flowering induction, shade avoidance response and seed germination. As a protein superfamily in plants, the family members and copy numbers vary in different plants. The PHYs of model plant bidopsis . There aomosomes . The chromophore .StPHY can be further subdivided into four subgroups homologous to PHYA/B/C/E progenitors. In the present study, we found that the structural domains in the StPHYs proteins are highly similar in type and arrangement. The distribution and functions of the structural domains of PHYs are consistent with the previous results, which can serve as a platform for protein interactions and a model for responding to changes in light conditions, oxygen levels and redox potentials [StPHYs gene, which were identical in number and type and aligned in the same way, showing a strong similarity. In this study, we also found differences in the positional distribution of the six structural domains of all members, which may account for the differences in the expression patterns of the members of the PHY gene family (The potato PHY gene family encompasses two physical types: photolabile type I and photostable type II. Concomitantly, according to the gene structure and sequence similarity, tentials . The N-ttentials . We idene family .A total of 16 cis-regulatory elements were identified within the putative promoter regions of genes encoding the potato PHYs gene family; they are responsive to phytohormones, abiotic stresses and photoresponses. PHY genes, as important light signaling receptor proteins in plants, play important roles in plant growth and development, morphogenesis, secondary metabolism and stress tolerance . PHYA isStPHYA and StPHYB varied in different organs, and the expression levels of individual genes displayed differences among different cultivars, these results showed that the expression patterns of StPHY genes were complex (StPHY members were induced by white light and red light; however, the increases induced by red light were the most significant (StPHYB/C/E were found to have the most obvious response to red light irradiation. According to the correlation analysis, the correlation coefficients between StPHYB/C and the α-solanine content were 0.75 and 0.68, while for α-chaconine they were 0.75 and 0.7. Thus, the GA content was significantly and positively correlated with the StPHYs gene expression. This result also indicated that the StPHYs gene play a more critical role in regulating GA biosynthesis. However, the detailed regulatory mechanism of StPHYs genes in response to light signaling and GA accumulation needs to be further investigated in future studies.In our work, the transcript abundance of  complex . After lnificant , showingStPHY genes were identified and their biological functions were clarified. Only the StPHYD subfamily member is missing from the StPHY gene family in potatoes. Although the functional structures are similar, there are obvious differences in expression patterns. Red light affects the content of GAs, and the photoreceptor protein StPHYs are involved in the regulation of GA content in potatoes. The availability of the StPHYs gene facilitates the exploration of the protein function of the gene and the mechanism of action of GA accumulation in response to different light signals. In addition, our research further refined the molecular regulatory mechanism of GAs. The study of photoregulation mechanisms in GA biosynthesis holds promise for targeted interventions of signaling pathways to reduce or even block them, providing a solution to the problem of greening and wasting of potatoes due to the accumulation of GAs.In this study, five