After collecting sputum, the strains from the tuberculosis analysis were isolated and confirmed. From a detailed examination of the mother and in accordance with the child's congenital tuberculosis, we confirmed the diagnosis of pelvic tuberculosis.We herein report a case of congenital tuberculosis. The infant was born at 27+5 weeks of gestation and was admitted to the hospital due to hypopnea after birth. Due to a poor response to treatment, we conducted pathogenic microorganism metagenomic analysis to assess the nucleotide sequences within the IVF treatment and pregnancy can exacerbate latent tuberculosis, especially in women from a family with a history of tuberculosis infections. We posit that the optimal way to prevent neonatal congenital tuberculosis in IVF-ET is to procure a detailed maternal medical or family history and to identify and treat maternal tuberculosis during IVF treatment. Mycobacterium tuberculosis (M. tuberculosis) infection, with an estimated 1.7 billion people (23%) infected worldwide; moreover, there are over 10 million new cases annually  and other assisted reproductive technologies (ARTs) over recent decades has allowed an increasing number of infertile women to conceive healthy babies, physical examination of the mother's own health is often ignored during the IVF treatment process, and this may conceal disease in the offspring. Among the existing publications on congenital tuberculosis, there are few reports of congenital tuberculosis infection in infants born from IVF. In this report, we describe a case of congenital tuberculosis in a premature infant born from IVF/ART and confirmed by macromicro (DNA) testing and positive acid-fast bacilli in sputum smears.Tuberculosis is a disease caused by n people 3% infectg et al. . Althoug9/L, neutrophils (NEU) at 42.50%, hemoglobin (HGB) at 155 g/L, platelets (Plt) at 208 × 109/L; a C-reactive protein (CRP) level < 0.5 mg/L; and a procalcitonin (PCT) of 0.41 ng/mL. Chest X-ray displayed lung changes and neonatal pneumonia in the premature infant, but plain abdominal X-ray revealed no abnormalities . The neonate's birthweight was 1,060 g, and his Apgar scores were 9-10-10. After birth, the boy was admitted to the NICU due to shallow breathing. Physical examination upon admission disclosed a body temperature of 36°C, a pulse rate of 144 beats/min, respiratory rate of 42 beats/min; and blood pressure of 66/44 mmHg. The child exhibited shallow breathing, grunting, and low breath sounds in both lungs; cardiac and abdominal examination, however, showed no abnormal signs. Initial examination at the time of hospitalization showed the following: blood test results disclosed white blood cells (WBCs) at 17.58 × 10malities . After a2) was maintained at 90–94%. During this period the child was breastfed and occasionally experienced abdominal distension, vomiting, and feeding intolerance. With a maximal feeding amount of 30 ml/Kg, body weight increased to 1,200 g, and the child's growth curve was at the 25th percentile.Mechanical ventilation was changed on the second day after hospitalization to non-invasive assisted ventilation, and we noted no apnea or periodic breathing under continuous non-invasive assisted ventilation over the next 14 days; transcutaneous oxygen saturation . However, blood cultures from two sites as well as sputum, stool, and urine culture did not divulge any bacterial growth. Chest X-ray displayed a thickening of lung markings and small flakes of fuzzy shadows, supporting a diagnosis of neonatal pneumonia. The abdominal intestine was also slightly distended, and intestinal dysfunction was considered  mode, the child manifested clinical symptoms such as dyspnea, obvious abdominal distension, and weakened bowel sounds. Thus, the boy's condition changed 2 weeks after being born prematurely; and although nosocomial infection was considered, the infection-related examination showed obvious abnormalities in his complete blood count -β-D-glucopyranose was 290.71 pg/mL ; B-ultrasonography revealed a small amount of fluid in the intestinal space. Chest X-ray examination displayed progressive aggravation of pneumonia, and the abdominal portion was slightly thickened and stiff on X-ray. According to the examination results, we recognized that the child possessed a serious infection, and meropenem and fluconazole were administered as anti-infectives.On day 18 after birth and under continuous mechanical ventilation, the boy's airway secretions increased, his sputum became thick, and erythematous, papulomacular, coalescent rash with indistinct borders . The blood test results showed 15.63 × 10th to 22nd days after birth. During this period, fever occurred for the first time; and even under continuous mechanical ventilation, his complexion remained cyanotic, and his TcSO2 was difficult to maintain at 90%. The difference in percutaneous oxygen saturation between the right upper extremity ductus arteriosus and the right lower extremity ductus arteriosus is larger than 10%. The partial pressure of oxygen (PO2) values from the blood gas analysis results on days 19–22 were 37.4 mmHg, 40.3 mmHg, 28.0 mmHg, and 34.6 mmHg, respectively, indicating that the child was hypoxemic. On day 22, blood test results showed that Plts decreased to 28 × 109/L and CRP increased to 134.8 mg/L. Chest X-ray examination on the same day showed large, dense opacities in the right upper lung field; small patchy opaque shadows in both lung fields; an air bronchogram sign; and a small amount of pleural effusion on the right side. We also noted bowel dilatation in the abdomen, and that the intestinal septum was slightly thickened, the left abdominal bowel was stiff, and the necrotizing enterocolitis (NEC) was aggravated  (The boy's condition further deteriorated from the 19gravated . The ven1 (100%) . As a re1 (100%) . TherefoThe child was, however, found to be in critical condition on day 24 after birth, and due to multiple organ-system dysfunctions and the family's withdrawal of treatment, the child was ultimately declared clinically dead.M. tuberculosis infection in the families of both parents, the mother had been infertile for many years; both of her pregnancies were via IVF/ET, each resulting in preterm births. The mother complained that although other infectious diseases were investigated during her IVF treatment, tuberculosis was not evaluated; and there was a history of persistent cough 1 month before delivery. We therefore recommend that a mother suspected of having tuberculosis be sent to a higher specialized hospital as soon as possible to complete the relevant examinations. The mother was ultimately diagnosed with pelvic tuberculosis at the Guangzhou Chest Hospital  and required conventional treatment.The neonate's father was 33 years old at the time of the boy's birth and in good health, and the boy's mother was 35 years old. There was a history of pregnancy, with the mother having undergone IVF-ET procedures and producing twins, and with premature rupture of membranes at 20 weeks of gestation. During the current pregnancy, the mother suffered from connective tissue disease and underwent premature rupture of membranes more than 6 days before delivery; and the child was born with clear amniotic fluid of approximately 30 ml. There had been numerous cases of M. tuberculosis complex. Upon receiving the metagenomic results, sputum was collected to detect acid-fast bacilli to confirm tuberculosis infection, and the mother was ultimately diagnosed with pelvic tuberculosis from the diagnosis of neonatal congenital tuberculosis.Our current patient was an IVF-ET neonate with congenital tuberculosis, a rare clinical manifestation. The neonate manifested hypopnea at his initial clinical appointment, was not febrile for 14 days after birth, and we noted no abnormality in the inspection results with regard to infection indicators; only the chest X-ray showed lung changes and neonatal pneumonia in this premature infant. Due to the unsatisfactory response to treatment and the worsening severity of his condition, serum metagenomic testing was considered in combination with evaluation for other uncommon pathogenic bacterial infections, and our results revealed Congenital tuberculosis is relatively rare in clinical practice, particularly in neonates born from IVF-ET. Furthermore, due to different clinical manifestations, misdiagnosis is more likely to occur and result in fatal consequences. While respiratory distress and fever are the principal symptoms of congenital tuberculosis , 6, shalvia inhalation of infected amniotic fluid at birth and the ingestion of infected substances . Written informed consent to participate in this study was provided by the participants' legal guardian/next of kin. Written informed consent was obtained from the minor(s)' legal guardian/next of kin for the publication of any potentially identifiable images or data included in this article.GZ and WL conceived the study. WL drafted the manuscript. LQ revised the manuscript. LY collected patient medical records. HZ supported laboratory data collection. All authors participated in the interpretation of the results and approved the version of the final manuscript.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


The mechanisms that underpin the species–area relationship (SAR) are crucial for both the development of biogeographic theory and the application of biodiversity conservation. Since its origin, the resource hypothesis, which proposes that rich resources in vast ecosystems will lower extinction rates and shape the SAR, has not been tested. The impossibility to quantify resources and extinction rates using plants and animals as research subjects, as well as the inability to rule out the influences of the area per se, habitat diversity, dispersal, and the historical background of biodiversity, make testing this hypothesis problematic. To address these challenges and test this hypothesis, two sets of microbial microcosm experimental systems with positive and negative correlated resources and volumes were created in this work. The results of 157 high-throughput sequencing monitoring sessions at 11 time points over 30 consecutive days showed that neither of the experimental groups with positive or negative correlations between total resources and microcosm volume had a significant SAR, and there were no negative correlations between extinction rates and resources. Therefore, in our microcosmic system, resources do not influence extinction rates or shape the SAR. Dispersal should be the principal mode of action if the resource theory is correct. The species–area relationship (SAR), which is the theoretical underpinning of island biogeography and an essential tool for biodiversity, as well as protected area designation, measures the positive link between species richness and habitat area ,2,3,4,5.According to the resource hypothesis of population ecology, population density rises as resource abundance rises, explaining the positive relationship between population density and habitat area ,13,14. AThe resource–density relationship has recently been supported by more and more evidences, although the density–richness relationship is still less clear-cut. The resource hypothesis, therefore, has yet to be put to the test in terms of shaping SAR. The resource hypothesis confronts three key obstacles in its validity. First, it is difficult to distinguish the role of resources from the role of area per se. SAR is shaped by both resources and area via extinction ,16, and Based on this, experiments must be planned to overcome the three problems raised above in order to test the resource hypothesis for shaping SAR and must investigate the following: (1) whether habitats with more resources have a higher population density of each species, and (2) whether habitats with more resources have lower extinction rates that shape the SAR, while satisfying the premise that larger habitats have more resources. To address these challenges, two sets of microcosm experiments  were established to test the resource hypothesis for shaping the SAR. The airtight, homogeneous, and consistent system of microbial origin ensures that influences such as dispersal, differences in biodiversity background , and habThere are four possible results from the experiments, which are as follows: (1) if both experimental groups have an SAR, the system’s SAR is completely determined by the area per se; (2) if neither experimental group has an SAR, extinction cannot independently shape the SAR, owing to resource disparities, and dispersion should be the more relevant process; (3) if group P has an SAR but group N does not, the SAR is only affected when the total resources is positively connected with area; (4) if group P does not have an SAR but group N does, neither resources nor area are the key variables that form SAR. Regardless of whether our results confirm the resource hypothesis, the outcomes of this work will offer fresh perspectives on SAR research thanks to the experimental approach used.3/NO2, and PO4 contents were also determined.The experimental design and the process of establishing the microcosm system for this study are shown in Pao cai  is a traditional fermented vegetable product that is high in microbial diversity and typically uses a variety of vegetables as raw materials. The vegetables are fermented in an anaerobic jar with a special pao cai brine.Raphanus sativus), 3.5 kg cabbage (Brassica oleracea), 0.2 kg chilli (Capsicum frutescens), 0.1 kg ginger , 0.1 kg Chinese-pepper (Zanthoxylum bungeanum), 0.25 kg rock sugar and 2 kg cold boiled water  and divided the solution into earthenware pao cai jars. After 7 days of natural fermentation at room temperature, the pao cai was filtered through sterile gauze to obtain 2000 mL of pao cai microbial seed solution, which was mixed well using a vortex shaker and set aside.We combined 3.5 kg white radish .The negative group was a microcosmic system in which the resource decreased with volume. First, eleven 50 mL, 100 mL, 150 mL, 300 mL, 500 mL, 700 mL and 1000 mL culture flasks were taken and 10 g, 7.0 g, 0.5 g, 5.0 g, 3.0 g, 1.5 g and 1.0 g of dry matter were added in that order and distilled water was added to the flasks .The above culture flasks were autoclaved at 121 °C for 30 min, cooled and 1 mL of pao cai microbial seed solution was added to each flask, replenished with sterile water until the flask was full and the flask was sealed without leaving any air inside. Each culture flask becomes a microcosm system. We placed each microcosm system in a constant temperature, light-proof incubator at 25 °C for 30 days.4-mg/L), and nitrate/nitrite content (NO3/NO2-mg/L) . Using a pH meter, the pH level of each sample was determined.Prior to the formation of the microcosm, one sample of well-mixed pao cai soup was used as a reference. After the microcosm system was constructed, samples were collected on days 1, 3, 6, 9, 12, 15, 18, 21, 24, 27, and 30, with one sample from each volume of the positive and negative groups taken at each sampling. Over the course of 30 days, 155 samples were collected at 11 different time points. After thoroughly mixing the microcosm system, 50 mL of culture fluid was taken as the sample, centrifuged at 8000 rpm for 10 min, and stored at −80 °C for precipitation. A segmented flow analyzer was used to determine the concentrations of total nitrogen (TN-mg/L), total phosphorus (TP-mg/L), orthophosphate content , according to the manufacturer’s protocols. For bacteria, we targeted the V3-V4 regions of the 16S ribosomal RNA (rRNA) gene using the 338F (5′-ACTCCTACGGGAGGCAGCAG-3′) and 806R (5′-GGACTACHVGGGTWTCTAAT-3′) primer pairs [Microbial DNA was extracted from the samples using the E.Z.N.A.er pairs . For funer pairs . PCRs wehttps://docs.qiime2.org/2019.1/, accessed on 5 March 2022). Briefly, FASTQ files that contained the raw data were imported into the QIIME format with the QIIME2 system, using the QIIME tools import program. Demultiplexed sequences from each sample were quality filtered and trimmed, de-noised, and merged, and then the chimeric sequences were identified and removed using the QIIME2 DADA2 plugin to obtain the feature table of amplicon sequence variants (ASVs) [Purified amplicons were pooled in equimolar amounts and paired-end sequenced (2 × 300) on an Illumina NovaSeq platform , according to standard protocols. The analysis was conducted by following the “Atacama soil microbiome tutorial” in Qiime2docs, along with customized program scripts (s (ASVs) ,28. An As (ASVs) .To determine if the microcosm system was set up in line with the pre-defined objectives, microbial diversity profiles were initially analyzed. Alpha and beta diversity were determined for each sample using the microeco and vegan packages in R 4.20 ,31. To sSecond, the correlation between resource abundance and population densities was examined to investigate whether greater resources could shape higher population densities; the correlation between extinction rates and volume, resources, and environmental factors was examined to investigate whether greater resource abundance could lessen extinction rates; and finally, SAR curves were built to investigate whether the resource hypothesis could influence SAR. The population density of each species of microorganism in high-throughput sequencing data is equivalent to the relative abundance of each species. The Kernel density estimate graph of the resources and relative abundance of each taxon was created using the ggplot2 package of R. The complete disappearance of an ASV is recorded as extinction. A mental test was conducted and a correlation graph for environmental factors and fungal species richness, bacterial species richness, and total extinction rate was created using the ggcor and ggplot2 packages . SAR curFinally, by utilizing microbial interaction network analysis, the influence of resources on microbial interactions in microcosmic systems was investigated. The mapping of microbial networks and the computation of network properties were carried out using the R package ggClusterNet .This study yielded a total of 13,159,681 16S rRNA gene sequences, which were then quality filtered, trimmed, and de-noised to yield 9781,102 high quality sequences, totaling 12,587 bacterial ASVs. A total of 11,386,587 ITS gene sequences were obtained and after quality filtering, trimming, and de-noising the sequences, a total of 8,678,214 high quality sequences were obtained, yielding a total of 10,433 fungal ASVs. The ASVs monitored covered 3 domains, 61 phyla and 1211 genera of microbial taxa.Firmicutes, Proteobacteria, Bacteroidetes, Fusobacteria, and Actinobacteria were the prominent bacterial phyla that co-occurred in the samples, while Ascomycota, Basidiomycota, Mortierellomycota, and Rozellomycota were the dominant fungal phyla that co-occurred in the samples  for each taxon. There was also no significant negative correlation between extinction rate and total resource, nor between extinction rate and volume. Resources did not change the structure or complexity of the microbial interaction network in the microcosm. These results imply that although the present study design has excluded many influences and ensured the premise that larger habitats have more resources, resources did not shape microbial SAR through extinction in the present experimental system. In contrast to cryoconite holes on less productive sites, Sommers et al. (2020) discovered that cryoconite holes on more productive locations accumulated more species with increasing hole area . ProductBased on insect population ecology research, some researchers believe that more resources will entice more insects to move into the area ,41. AccoIn essence, biodiversity is determined by only three processes, speciation, dispersal, and extinction . The SARThe future of SAR, and indeed biodiversity research in general, should return to the three key processes, for which microbial microcosm experiments are uniquely suited. Firstly, microcosm experiments enable continuous monitoring of temporal changes in microbial diversity. Studies have found that Escherichia coli can reproduce one generation every 20–30 min. In the 30-day microcosm experiment of this study, the microorganisms may have reproduced for more than 2000 generations. From the perspective of humans, the 2000 generations have spanned 50,000 years of changes . Only microbial microcosmic systems are capable of conducting research with such a large generation span. Secondly, the microcosmic system is able to independently verify the individual factors that influence microbial diversity. The dilemma with current research on SAR mechanisms is that the factors that influence species richness in natural habitats are often indistinguishable from each other . For exaBased on the results, we believe that resources cannot independently shape the SAR through extinction; however, we cannot rule out the possibility that resources shape the SAR through factors such as dispersal. We call for future research to further validate this relationship from a dispersal perspective.


This study aimed to determine trajectories of depressive symptoms among older adults in England, overall and for those with hip fracture. The study aimed to explore the differential characteristics of each trajectory identified.n = 7 050), including a hip fracture subgroup (n = 384), from the English Longitudinal Study of Ageing. Latent class growth mixture modeling was completed. Depressive symptom prevalence was estimated at baseline. Chi-square tests were completed to compare baseline characteristics across trajectories.Analysis of adults aged 60 years or more (p < .001).Three trajectories of depressive symptoms  were identified overall and for those with hip fracture. The moderate-severe trajectory comprised 13.7% and 7% of participants for overall and hip fracture populations, respectively. The proportion of participants with depressive symptoms in the moderate-severe trajectory was 65.4% and 85.2% for overall and hip fracture populations, respectively. Depressive symptoms were stable over time, with a weak trend toward increasing severity for the moderate-severe symptom trajectory. Participants in the moderate-severe symptom trajectory were older, more likely to be female, live alone, and had worse health measures than other trajectories (Older adults, and those with hip fracture, follow one of the 3 trajectories of depressive symptoms that are broadly stable over time. Depressive symptoms’ prevalence was higher for those with hip fracture and, when present, the symptoms were more severe than the overall population. Results suggest a role of factors including age, gender, and marital status in depressive symptom trajectories. Some stUnanticipated health care events such as hip fracture may negatively influence trajectories of depressive symptoms in older adults . CristanThis study aims to determine the trajectories of depressive symptoms among older adults in England over a 17-year period and whether these trajectories vary for those with hip fracture. This study will also explore the differential characteristics of each trajectory identified.Participant data for this study were obtained from the English Longitudinal Study of Ageing (ELSA) data set, a nationally representative longitudinal study of community-dwelling adults aged 50 years and older . Data onThe analysis cohort for this study (ELSA Waves 1–9) were over the age of 60 years in the year they entered the ELSA sample, with recorded 8-item Centre for Epidemiological Studies-Depression (CES-D) data in at least 2 waves , and CESDepressive symptoms were measured using the 8-item CES-D scale . The scaAlcohol consumption, social networks, Control Autonomy Self-realization Pleasure-19 (CASP-19) , and theAnalyses were completed for the entire population and for the subgroup of older adults reporting a hip fracture at any wave. This approach was taken due to the potential of an unexpected health care event such as a hip fracture leading to different trajectories of depressive symptoms over time . BaselinGroup-based trajectory modeling (GBTM), a type of latent class growth mixture modeling application, group individuals by estimating latent trajectories in a population . GBTM waWe tested a series of models using either 2, 3, 4, or 5 groups and combined different polynomial shapes; 0 = intercept, 1 = linear, 2 = quadratic, 3 = cubic. To determine the model of best fit and the optimal number of trajectories, we adopted several model fit indices; the Bayesian information criteria (BIC) , entropyFor the overall sample, data were used from the individual’s entry wave. For the hip fracture subgroup, data were used from the wave which included the first reported hip fracture (fracture wave). The proportion of patients with depressive symptoms (numerator as the number of participants whose CES-D ≥ 4 and denominator as the number of participants in each group) at baseline (entry wave) was calculated overall and for each trajectory. Differences between trajectory characteristics were assessed with the Chi-square test . Chi-squThe trajectory analysis method employed automatically imputes missing data for individuals with depressive symptoms measured at least twice. Overall, 7 050 patients over the age of 60 were included in the ELSA data set in England between 2002 and 2019 . Of thesCharacteristics of the study cohort are detailed in n = 2 726), “mild” (n = 3 357), and “moderate-severe” (n = 967) symptom trajectories .There were statistically significant differences between the 3 trajectories for the prevalence of depressive symptoms, age, sex, ethnicity, marital status, activities of daily living, instrumental activities of daily living, total number of comorbidities, total number of mobility limitations, number of times fallen, CASP-19 score, self-rated general health, life satisfaction score, if health limited ability to work, pain, physical activity level, having a longstanding illness, comorbidities of the musculoskeletal, respiratory, circulatory and metabolic systems, falls history, alcohol consumption, social network, smoking status, if they fractured their hip or had depression or manic depression (n = 138), “mild” (n = 219), and “moderate-severe” (n = 27) symptoms . In the overall population, there were 3.6%, 6.2%, and 8.2% of patients with hip fracture in the no-, mild-, and moderate-severe symptom trajectories, respectively. Overall, 384 (5.5%) patients in the ELSA data set suffered a hip fracture between 2002 and 2019. Of these, 87 (22.7%) patients experienced depressive symptoms at the fracture wave. A 3-group model in the order of 0, 0, 0 was the most appropriate, when the best values of the model selection criteria were considered together. The summaries of the model selection by model and by the groups in the chosen model as well as the posterior probabilities for the selected model are presented in symptoms . Depresssymptoms . The chap < .001). There was no significant difference between the 3 trajectories for ethnicity, age, sex, BMI, activities of daily living, instrumental activities of daily living, total number of comorbidities, falls history, physical activity level, comorbidities of the circulatory, respiratory, musculoskeletal, and metabolic systems, mental disorders and neoplasms, alcohol consumption, smoking status, social networks, and whether they had diagnosis of depression or manic depression , and moderate or increasing and persistent . The resAmong older adults after hip fracture, Cristancho et al. identified 3 trajectories “resilient,” “distressed,” and “depressed” . In contA higher median CES-D score was observed for the moderate-severe trajectory for the hip fracture subgroup (6.5 [IQR: 6.0–7.5]) compared with the moderate-severe trajectory for the overall population (4.0 [IQR: 3.0–6.0]). This may suggest greater symptom severity among those with a history of hip fracture. Although the sample size for the moderate-severe symptom trajectory in the hip fracture group was small, the median CES-D score and its IQR were more compact. This suggests there was less variation in the population of the moderate-severe symptom trajectory among patients with hip fracture than the overall sample. This is not surprising, as the hip fracture group is expected to be more homogeneous than the overall, larger sample.There were differences in the characteristics of older adults within each trajectory, overall, and for those with hip fracture. For example, individuals in the trajectory with moderate-severe symptoms were older, more likely to be female, less likely to be married or in a civil partnership, and exhibited worse health and quality of life outcomes. These results are comparable to Musliner et al. who reported in the higher symptom trajectories, individuals were more likely to be female, were smokers, and have poor self-rated general health . For aduSimilar to the study by Liang et al., the current study noted those alone exhibited higher depressive symptoms than those married both overall and for those with hip fracture . In contThe extent to which participants in previous studies were in receipt of pharmacological and/or nonpharmacological management for depressive symptoms over time is poorly described. The effectiveness of these management approaches in the general adult population is well established  and are There are limitations to this study. First, the ELSA sample comprises predominantly White community-dwelling individuals limiting generalizability of results to non-White and residential/nursing care populations. Second, data were missing for several variables that were not collected across all waves. We assessed the sensitivity of our findings to missingness by conducting the analyses using data from the first 5 waves only and then by excluding missing depressive symptoms. We found the results comparable to the main analysis. Third, there is potential selection bias as the year individuals suffered their hip fracture was not available and may relate to the trajectories of depressive symptoms. This also meant we were unable to see the level of change in depressive symptoms after hip fracture. Fourth, there are no gold standards for sample size for latent class analyses. Sample sizes as small as 30 have been shown to produce valid results for simple latent class modeling with distinct classes, whereas sample sizes from 300 to 1 000 have been suggested for models with multiple indicators and classes. The main concerns regarding inadequate sample size are unpowered models which cannot detect classes, inaccurate solutions produced, and small but relevant classes being missed . HoweverOlder adults, and those with hip fracture, follow one of the 3 trajectories of depressive symptoms that are broadly stable over time. This may suggest it is never too late to target depressive symptoms for these patients as symptoms may persist in the longer term. Only one of the 3 trajectories had a median CES-D score, which met the threshold for depressive symptoms. Depressive symptoms’ prevalence was higher for those with hip fracture and the distribution of depressive symptoms consistently over time is shifted toward the moderate-severe group for those in the hip fracture subgroup when compared with the overall population. Results suggest a role of factors including age, gender, and marital status in depressive symptom trajectories both overall and for those with hip fracture which should be explored in future research.glac182_suppl_Supplementary_MaterialClick here for additional data file.


The objective of the present study is to evaluate the association of red blood cell distribution width with acute kidney injury in sepsis. This is a retrospective study of 849 critically ill patients with sepsis in intensive care unit. Demographic data, renal function, inflammation, complete blood count, and acid-base parameters were compared between acute kidney injury and non-acute kidney injury groups. Therefore, a multivariate analysis was performed to observe independent predictive factors.2/FiO2 ratio in the non-acute kidney injury group. In addition, there was an independent association of comorbidity-chronic kidney disease , urea  and RDW  with acute kidney injury in sepsis patients. Comparatively, higher levels of C-reactive protein, lactate, red blood cell distribution width, and Simplified Acute Physiology Score 3 were found in the acute kidney injury group. The study showed a higher frequency of women, hemoglobin (Hgb) concentration, platelets, bicarbonate and PaO As an elective risk factor, red blood cell distribution width was independently associated with sepsis-related acute kidney injury. Thus, red blood cell distribution width acts like a predictive factor for sepsis-induced acute kidney injury in intensive care unit admission. Sepsis is the most important cause of AKI in critically ill patients with high mortality in the intensive care unit (ICU).-6 In addition, AKI in sepsis may also be distinguished from non-sepsis AKI by some specific features and poor prognosis.In line with this, sepsis-associated AKI is complex and multifactorial. It includes endothelial and tubule changes, infiltration of inflammatory cells, and intrarenal hemodynamic changes with intraglomerular thrombosis. In some cases, changes in RDW indicates blood-associated abnormalities or organ/systems dysfunction and acts as a predictor of outcomes in critically ill patients.-12Considering the hematological context, red cell distribution width (RDW) measures the amount of red blood cell variation in volume and size. The RDW is defined as the standard deviation of erythrocyte size divided by the mean corpuscular volume.-10 Further, RDW can integrate various pathophysiological mechanisms associated with sepsis.,9,11-13 Recently, researchers have reported an independent association of RDW and the risk of adverse outcomes in patients with sepsis.,11 However, no evidence of the RDW association with sepsis-induced AKI was found. Therefore, this study analyzed the relationship of RDW and sepsis-induced AKI.In addition, RDW is a readily available biomarker related to decreased hemoglobin levels by inflammatory cytokines and impairing iron mobilization.To analyze whether red blood cell distribution width could act as a predictor of acute kidney injury in patients with sepsis.Hospital Israelita Albert Einstein (HIAE), São Paulo, Brazil, from January 1 to December 31, 2017, were retrospectively studied. First, the study observed that the patients were screened by quick Sequential [Sepsis-related Organ] Failure Assessment (qSOFA) for early suspected sepsis in the emergency department or bedside in the ward. A qSOFA ≥2 was considered altered with the following variables: Glasgow coma score <15, respiratory rate ≥22 breaths/minute, systolic blood pressure ≤100mmHg. Afterward, the patient was referred to the ICU.Medical records of patients with sepsis who were admitted to any unit of the Intensive Care Center of Inclusion criteria were individuals over 18 years of age with a diagnosis of sepsis admitted to an ICU at HIAE. Patients with hematologic, advanced oncological disease, liver failure, chronic infection by HIV, hepatitis B and C, chronic kidney disease (CKD) stages 4 or 5, and patients who died within the first 24 hours of admission to the ICU were excluded.2/FiO2) ratio, and Simplified Acute Physiology Score 3 (SAPS 3) prognostic index. Outcomes such as AKI, need for Renal Replacement Therapy (RRT), red cell transfusion, mechanical ventilation, vasopressor drugs, and mortality were computed. Norepinephrine was the primary vasopressor used when mean arterial pressure was maintained <65mmHg after volume resuscitation. Continuous venovenous hemodiafiltration was used in patients with hemodynamic instability, and intermittent hemodialysis was used in stable hemodynamic patients who required RRT during the ICU period.The variables analyzed were age, sex, Hgb concentration, blood count parameters such as mean corpuscular volume, RDW, platelet, white blood cell count, serum levels of urea, creatinine, sodium, potassium, total bilirubin, C-reactive protein (CRP), arterial blood gas parameters, arterial lactate, arterial oxygen pressure/inspired oxygen fraction . The study was conducted under the ethical standards of the responsible committee on human experimentation . Informed consent was waived, and researchers analyzed only not identified data.Acute kidney injury was defined and stratified according to the Kidney Disease Improving Global Outcomes (KDIGO) criteria, if there was an increase of 0.3mg/dL in 48 hours, or a 1.5-fold increase in serum creatinine in seven days, or diuresis lower than <0.5mL/kg/hour during six hours from ICU admission.2 test for frequencies was used. The Kolmogorov-Smirnov test was performed to find the normal distribution of continuous data. Afterward, the comparisons between the groups were made by the Student’s t-test for data with normal distribution and Mann-Whitney for non-parametric data. The Pearson correlation between two continuous variables was also performed, in addition to the binary logistic regression with the backward stepwise method for admission data using AKI as a variable response. The variables in the multivariate model were included when reached p<0.09. Regression data were expressed as odds ratios (OR) and 95% confidence intervals (95%CI). Moreover, the association by regression of ICU admission markers with the need for RRT and mortality was also determined. For that analysis, the marker variables from ICU admission that were independently significant with AKI were used. The statistical level p<0.05 was considered significant. Statistical analyses were performed in SPSS version 22  and ExcelTM 16.0 .Categorical variables are presented as frequencies and percentages. Numerical data were described as mean ± standard deviation. In a comparative analysis, the χAfter exclusion criteria , the stuA positive correlation was found between simplified acute physiology score 3 and red blood cell distribution width in 849 patients with sepsis (p<0.001).Furthermore, this study found that, among the total of sepsis patients, 294 had AKI diagnosis by KDIGO criteria. A total of 160 patients were in stage 1, 73 in stage 2, and 61 in stage 3, considering the KDIGO criteria. Thereafter, patients with sepsis were divided into two groups: AKI and without AKI (non-AKI) to perform several comparisons and associations.diabetes mellitus (DM), hypertension, and CKD) compared to the non-AKI Group. Lower indices for mean arterial pressure, Hgb concentration, platelets, pH, pCO2, arterial bicarbonate, and PaO2/FiO2 ratio in AKI Group were found in comparison to non-AKI. Moreover, the AKI Group had higher mean age, and higher levels of creatinine, urea, total bilirubin, RDW  at intensive care unit admission was found in sepsis-induced AKI Group compared to non-AKI Group (p<0.001).In sequence with this analysis, Subsequently, considering the overall outcomes observed during ICU stay of sepsis patients (n=849), it was highlighted: 165 patients (19.4%) required mechanical ventilation in 3.9±2.4 days; 151 patients (17.8%) required red cell transfusion in 8.1±6.4 days; 148 patients (17.4%) required vasopressor drug in 5.1±2.9 days; 73 patients (8.6%) evolved with mortality in 14.2±5.4 days; and 34 patients (4%) required RRT in 9.6±6.9 days after admission to the ICU. An association of RRT requirement with urea  and RDW  on ICU admission was also identified. A total of 18 patients (52.9%) in RRT required continuous venovenous hemodiafiltration. The study also found that mortality within 28 days of ICU period had an association with levels of RDW  and urea  of admission to ICU.Finally, a subgroup of patients without CKD (n=759) was analyzed to assess and rule out possible CKD interference. This subgroup of patients without CKD but with sepsis-associated AKI (n=219) had higher levels of RDW . There was also an independent association of RDW with sepsis-induced AKI in patients without CKD .The main finding of the present study is that RDW is an independent predictor associated with AKI in critically ill patients with sepsis. In addition, the study identified that the highest RDW on admission was associated with the need for RRT during the ICU period.,18 Therefore, the relevance of the current study is corroborated by recent studies.,19The patients with sepsis-associated AKI had lower mean arterial pressure and arterial bicarbonate, but with elevated arterial lactate at admission, which symbolizes a situation according to low perfusion and organ dysfunction that often occurs in sepsis. When correlations were analyzed in patients with sepsis, negative associations of bicarbonate with both lactate and creatinine were identified. In addition, a positive correlation of RDW value with variables of poor renal function (urea and creatinine) was found.Sepsis is the most important cause of AKI in the ICU and it is estimated that 15% to 20% of patients with sepsis-associated AKI need RRT.,16,19,20 Thus, the present study could define and form groups of patients with and without AKI. The group of patients who developed sepsis-associated AKI had lower PaO2/FiO2 ratio, but higher levels of urea, creatinine, arterial lactate, RDW, and SAPS 3 prognostic index. All these clinical parameters indicate kidney dysfunction, decreased oxygenation, and lower tissue perfusion. Thus, there is more severity and poorer prognosis in patients with AKI associated with sepsis. On the one hand, this study group has recently demonstrated that critically ill patients with COVID-19-related AKI had a lower PaO2/FiO2 ratio too. Indeed, lung dysfunction from AKI is associated with poorer outcomes. On the other hand, Loveday et al. reported the addition of RDW to APACHE III increased the prognostic value in critically ill patients. Moreover, serum creatinine as a marker of renal function is a variable of the SAPS 3 prognostic index, reflecting greater levels of this prognostic index in patients with AKI.,15 Furthermore, the current study found a positive correlation between the prognostic index-SAPS 3 and RDW. This indicates a relationship between RDW and prognostic index for critically ill patients. However, the present results corroborate with the results of other studies.The diagnosis of AKI is currently based on increased serum creatinine concentration and/or decreased urine output. On the other hand, arterial lactate did not present an independent association with sepsis-induced AKI in the current study. Even so, this study found that the levels of bicarbonate on admission to the ICU were lower in the group that developed AKI. In addition, a multivariate analysis showed that higher levels of arterial bicarbonate have a protective effect on developing AKI. These findings agree with the results of Jung et al., who observed that low levels of serum bicarbonate were associated with a higher incidence of AKI and prolonged ICU stay after cardiac surgery.Moreover, the results of the present study were different from the results of Radovic et al., which observed levels of arterial lactate as an independent predictor of AKI but in patients undergoing cardiac surgery.,8,12 Red cell distribution width is reported as a marker and predictive value in many clinical settings. Changes and disorders frequently seen in sepsis such as ineffective erythropoiesis or increased destruction of red blood cells cause anemia and consequently greater heterogeneity in the size of these cells and higher RDW.,9,10 Moreover, previous studies showed that RDW is associated with AKI in patients with traumatic brain injury, sepsis in pediatric patients, after cardiac surgery, and those with coronary disease.-28The RDW is routinely reported as part of a complete blood cell count and expresses variation in the size of circulating erythrocytes.2/FiO2 ratio, platelets, and Hgb concentration in patients with sepsis and AKI,,8,17 it can be assumed that when inflammation occurs, it reduces iron metabolism, and bone marrow function is inhibited, thus, the erythrocyte proliferation and maturation are inhibited,,30 leading to increased RDW values and anemia, while inflammatory dysregulation can reduce tissue perfusion and thus cause AKI.The present study confirmed the results found by other researchers. Moreover, as there were higher levels of CRP, arterial lactate, and RDW, in parallel with lower arterial bicarbonate, PaOThis study also found that RDW levels were higher in patients who developed AKI and showed that RDW at admission to ICU had an independent association with AKI in multivariate analysis. Interestingly, each 1% of the increase in the RDW value measured was independently associated with important risk (16%) of sepsis-induced AKI. Furthermore, each increase of 1mg/dL in the value of urea (higher than the cut-off of 50mg/dL) at admission was independently associated with a risk of 4% more of sepsis-associated AKI. The current study also found that female sex, and higher serum bicarbonate levels on admission to the ICU were associated with protection against sepsis-induced AKI. Even so, the present results showed a relationship between RDW and sepsis-associated AKI also in the subgroup of patients without CKD. Thus, eliminating the interference of CKD in this relationship.Furthermore, it was also detected that the patients with CKD in stages 1-3, had 3.5 times more likely to develop AKI associated with sepsis independently. Other researchers reported a bilateral interaction of CKD with AKI. Chronic kidney disease predisposes to AKI. On the other hand, severe AKI could worsen the progression of CKD. Fernandez et al. reported higher values of RDW as a marker of severity in patients discharged from the ICU.Subsequently, the association of variables at ICU admission independently related to AKI with outcomes within 28 days was also analyzed. Each increase of 1mg/dL in the value of urea higher than the cut-off of 50mg/dL at ICU admission was associated with an increased risk (2.3%) of need for RRT. In turn, each 1% increase in the RDW value measured was associated with the risk (18%) of need for RRT. Also, there was association between mortality within 28 days of ICU stay and both urea and RDW value at ICU admission. Thus, each 1% increase in the measured RDW value was independently associated with 37% likely of mortality at 28 days of ICU stay. Thereby, these results are in robust compliance with other studies. In fact, Oh et al. reported that RDW is a predictor for all-cause mortality in AKI patients on RRT treatment in the ICU.Although intriguing, the present study has some potential limitations. First, the sample evaluated is from a retrospective cohort study. Second, it was in a single-center, there was no intervention by the researchers or sequential analysis of RDW, and there was no pre-specified hypothesis. Finally, given the large number of potential predictors evaluated and the initial lack of selection guided by hypotheses of variables, the possibility of confounding bias cannot be ruled out.Despite these limitations, one of the main strengths of this study is that it is based on routine blood tests for critically ill patients on admission to the ICU and is inexpensive. Thus, a strong and independent association of RDW with sepsis-induced AKI was found, reflecting severity and organ dysfunction. However, the present study showed that the RDW on ICU admission acts as a predictor of AKI associated with sepsis.This study concluded that red cell distribution width can be used as a cost-effective marker for sepsis-associated acute kidney injury. Thus, further studies should be conducted to analyze the true predictive value of red blood cell distribution width in sepsis-induced acute kidney injury.


Early detection of synchronous colorectal peritoneal metastases (CPMs) is difficult due to the absence of typical symptoms and the low accuracy of imaging examinations. Increasing the knowledge of the risk factors for synchronous CPM may be essential for early diagnosis and improving their management. This study aimed to identify the risk factors for synchronous CPM.The study was registered at PROSPERO (CRD42020198548). The PubMed, Embase and Cochrane Library databases were searched for studies comparing the clinicopathological and molecular features between patients with or without synchronous CPM. The pooled data were assessed by a random-effects model.Twenty-five studies were included. A synchronous CPM was positively associated with female sex , PROK1/PROKR2-positivity , right-sided colon cancer , poorly differentiated grade , BRAF mutation , mucinous adenocarcinoma , signet-ring cell carcinoma , N1-2 , T4  and elevated serum CA19-9 .These evidence-based risk factors are indicators that could predict the presence of synchronous CPMs and can improve their management.www.crd.york.ac.uk/prospero, identifier: CRD42020198548. Despite the recent improvements in cancer research, colorectal cancer (CRC) has the second highest mortality in both men and women worldwide . An impoThe early detection of synchronous colorectal peritoneal metastasis (CPM) is currently difficult due to the absence of typical symptoms and the low accuracy of noninvasive imaging examinations for nodules smaller than 5 mm . In factSome studies have been previously conducted in order to identify the risk factors associated with synchronous CPM, but they have had heterogeneous outcomes, such as the location of the primary tumour , 10 and Therefore, a comprehensive understanding of the clinicopathological and molecular characteristics of CPM may be necessary for early diagnosis and may help to improve the management of patients who are at high risk of synchronous CPM. A systematic review and meta-analysis of all studies comparing sex, tumour invasion depth, lymph node metastasis, differentiation, location of primary tumour, histological results, and the serum levels of CA19-9, PROK1/PROKR2, BRAF, KRAS, NRAS, PIK3CA and MSI-H/dMMR between synchronous pmCRC and nonpmCRC patients was undertaken.This systematic review and meta-analysis adhered to the recommendations of the Preferred Reporting Items of Systematic Reviews and Meta-analysis (PRISMA) statement . The PRIwww.crd.york.ac.uk/prospero). Number CRD42020198548.This study was registered at PROSPERO  were eligible for inclusion. The included studies met the recognized diagnostic criteria as follows: primary colorectal tumours; the primary tumour’s pathological diagnosis was confirmed; and the patient’s synchronous PM was confirmed by an imaging diagnosis before surgery, an intraoperative exploration or by a histopathological examination.The exclusion criteria were as follows: (1) case reports, review articles and animal studies; (2) non-English publications; (3) studies that were not related to CRC or PM; (4) metachronous PM; (5) no analysis of the risk factors; (6) no comparator group; (7) no relevant data, including articles published only in abstract form as well as studies without complete data or an inability to construct a 2×2 contingency table from the present data; (8) mixed primary tumours; (9) a nonstandardized histological type; and (10) synchronous CPM was not clearly or correctly defined.We selected relevant studies by searching PubMed, Embase and the Cochrane CENTRAL Register of Controlled Trials. The following combined terms were used in the search:  AND . The latest search was implemented on July 14, 2020, and there was not limit to the earliest date of publication.Two independent authors (Y Zhang and X Qin) checked the title and abstract of each study, and the studies that satisfied the potential eligibility were obtained for further full-text assessment. Disagreements were resolved by discussion with the senior authors (Huaiming Wang or Hui Wang) until a consensus was achieved.By using standardized forms, two authors (Y Zhang and X Qin) independently extracted the data from each eligible study. The authors resolved any disagreements by discussion with the senior authors (Huaiming Wang or Hui Wang). The following data were extracted from each eligible study: author, year of publication, country where the study was conducted, setting of the centre, type of study, enrolment interval, number of primary CRC patients with or without synchronous PM and the clinicopathological and molecular characteristics. In addition, the Newcastle–Ottawa Scale score (N-O score) was also calculated and extracted for all of the eligible studies.2-based Cochran Q test was used to assess heterogeneity among the studies, in which P < 0.1 indicates the presence of heterogeneity  and Stata (version 12.0) for all statistical analyses. All of the pooled outcomes were determined using a random effects model (DerSimonian–Laird method). In the pooled analyses of the associations between the clinicopathological-molecular characteristics and synchronous PM, the effect sizes were calculated as the odds ratios (ORs) with 95% confidence intervals (CIs). The χogeneity . We alsoogeneity . For sigogeneity .The qualities of the included studies were assessed using the Newcastle–Ottawa Scale , in whicThe initial search yielded a total of 9470 studies. After removal of duplicates, a total of 7659 studies were screened by analysing their titles and abstracts, and 7435 studies were removed because they met one or more of the exclusion criteria. The remaining 224 studies were then assessed for eligibility by full-text examination, and a further 199 were excluded due to ineligibility. The reasons for exclusion were recorded. Finally, 25 studies were included in the final analysis Figure 1Among the 25 included studies, 7 had a multicentre setting, and 18 had a single centre design. Five of the included studies were prospectively performed; the remaining twenty were retrospective. All included studies were considered high quality (N-O score ≥ 6). Complete characteristics of the included studies are available in Six clinicopathological and molecular factors could not be included in the quantitative synthesis because they had only a single study in their subgroup, or their methodology did not permit for the pooling of the data. The six factors were serum CEA , serum CFinally, 21 studies on 13 factors were included in the quantitative synthesis through the meta-analysis: 7 studies on sex, 4 studies on the tumour invasion depth, 3 studies on lymph node metastasis, 5 studies on the differentiation, 6 studies on the primary tumour site, 7 studies on the histological findings, 2 studies on serum CA19-9, 2 studies on PROK1/PROKR2, 9 studies on BRAF, 6 studies on KRAS, 2 studies on NRAS, 2 studies on PIK3CA and 4 studies on the MSI-H/dMMR status.Seven studies , 30, 38 Four studies , 22, 38 Three studies , 22, 38 Five studies , 38, 40 Six studies , 22, 38 Five studies , 22, 38 Six studies , 38, 40,Seven studies , 38, 40,Three studies , 23, 35,Levels of up to 37.0 µ/ml were taken as the upper cut-off values for the Serum CA19-9 reference ranges. Two studies , 36, whiTwo studies , 34, whiNine studies , 30, 32 Six studies , 25, 30,Two studies , 25, whiTwo studies , 30, whiFour studies –13, 28, No significant publication bias was found according to the visual inspection of the funnel plot and Begg’s test S6.This study has provided an extensive analysis for the association between synchronous CPM and its clinicopathological-molecular features. We found that synchronous CPM was positively associated with female sex, PROK1/PROKR2 positivity, a right-sided colon cancer location, a poorly differentiated grade, BRAF mutations, mucinous adenocarcinoma, signet-ring cell carcinoma, N1-2, T4 and an elevated serum CA19-9 . However, synchronous CPM was not associated with KRAS, NRAS, or PIK3CA mutations or MSI-H/dMMR.Some studies have previously defined the degree of risk of developing colorectal peritoneal carcinomatosis , 42. A hBased on the hypothesis that phenotype and the subsequent clinical behaviour of CPM are driven by underlying biological mechanisms, studies that investigate disease biology will contribute to more precise identification of the suitable patients and for the guidance of therapy. This is one of the critical future research targets in CPM research. The potential mechanisms of the risk factors that are positively associated with synchronous CPM are discussed below. Due to a longer asymptomatic period, right-sided colon tumours are usually larger in diameter when they are diagnosed than left-sided colon tumours are. Larger neoplasms infiltrate the surface of the serosa over a larger area, which may lead to increased abscission of cancer cells into the peritoneal cavity. In addition, typical genetic differences between right-sided and left-sided colon tumours have been found, such as the BRAF status, and these genotypes may bring about a phenotype with a different probability of being associated with synchronous CPM . SeveralThere are some limitations in this study. First, non-English studies were excluded causing a language bias. Second, the risk associated with T4a vs. T4b stage was not analysed because there are no such detailed data. Finally, the number of included studies regarding CA19-9, PROK1/PROKR2, NRAS and PIK3CA was small, which may have limited its statistical power. We look forwards to conducting further studies.To our knowledge, this is the first meta-analysis to reveal the clinicopathological and molecular features of synchronous CPM. These evidence-based risk factors are conducive to strengthening the patient management and selecting the optimal therapeutic strategy.The datasets used and/or analysed during the current study are available from the corresponding author on reasonable request.HW designed the study. YZ, XQ, HMW, and HW performed the study selection and data extraction. YZ and RL performed the statistical analysis and data interpretation. YZ and XQ wrote and edited the manuscript. HL revised this manuscript. All authors read and approved the final draft.This study was sponsored by the National Natural Science Foundation of China (grant No. 82103084) and the Sun Yat-sen University Clinical Research 5010 Program (grant No. 2019021). The funding body had no role in the design of the study and collection, analysis, and interpretation of data and in writing the manuscript.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


This retrospective study aimed to describe a pediatric canine population presumptively diagnosed with acute viral gastroenteritis and to identify potential prognostic factors that influence hospitalization time (HT) and mortality.Acute viral gastroenteritis is one of the main causes of hospitalization in dogs during the 1Canine patients up to 12 months of age diagnosed with presumptive acute viral gastroenteritis were searched retrospectively from two veterinary hospitals during a 5-year period (2015–2020). Information regarding patient signalment, prophylactic care, clinical signs, blood test results, presence of systemic inflammatory response syndrome, and additional treatments were recorded to analyze their association with HT and mortality. Only dogs with a complete medical record until death or discharge were included in the study.Ninety-four dogs were identified: 76 dogs (80.9%) survived with a median HT of 5 days (range: 2–16 days) and 18 dogs (19.1%) died with a median HT of 3½ days (range: 1–8 days) after admission. The presence of fever and fresh frozen plasma (FFP) administration was significantly associated with a lower survival rate (p = 0.021 and p = 0.037) in the multivariate analysis. Among survivors, incomplete primo-vaccination, the presence of hematochezia, and FFP administration were considered independent predictors of time to clinical recovery , being associated with higher HT.The presence of fever and FFP administration was significantly associated with a lower survival rate. An inadequate primo-vaccination status prior to admission, hematochezia, and FFP administration was associated with longer HT in surviving patients. Further studies are needed to confirm the present results. Canine viral gastroenteritis represents a group of diseases characterized by viral-induced gastrointestinal inflammation . AlthougCanine parvovirus is the most frequently diagnosed pathogen, causing the destruction of rapidly dividing cells present in intestinal crypts and lymphoid organs, leading to the manifestation of acute-onset signs of vomiting, diarrhea, lethargy, and anorexia . AnimalsTo prevent viral infection, at-risk puppies must be kept protected from possible exposure until they complete a course of vaccinations from 6–8 weeks through 16 weeks of age or older, according to the latest World Small Animal Veterinary Association vaccination guidelines [The standard treatment for viral gastroenteritis includes fluid therapy, broad-spectrum antibiotics, antiemetics, gastroprotective drugs, and interventional nutrition , 7. FresThe prognosis of dogs with viral gastroenteritis has already been associated with the severity of clinical signs and laboratory abnormalities at the time of diagnosis, the presence of comorbidities, and the therapeutic approach elected , but furThis retrospective study aimed to describe a pediatric canine population presumptively diagnosed with acute viral gastroenteritis and to identify potential prognostic factors that influence hospitalization time (HT) and mortality.Because of the retrospective nature of this study and the use of diagnostic data collected as a part of routine clinical procedures, the need for ethical approval was waived.The archived medical records (from January 2015 to December 2020) of two referral veterinary hospitals in the northern region of Portugal were searched for juvenile dogs diagnosed and treated for acute gastroenteritis presumed to be viral in origin.Inclusion criteria were patients diagnosed with presumptive acute viral gastroenteritis up to 12 months of age and with a complete medical record until the time of discharge or death. Dogs were excluded if the cause of gastroenteritis was related to toxin ingestion, gastrointestinal foreign bodies, sudden change in diet, recent medication, or even infectious agents other than viruses . Moreover, patients were excluded if they had an incomplete medical history or an inadequate follow-up due to insufficient data regarding their clinical case or outcome.For each dog, patient information regarding signalment, such as age, sex, weight, breed and size, prophylactic care, including deworming and vaccination status, specific clinical signs, in particular hematochezia and fever, blood test results, namely, nadir neutrophil count and serum albumin concentration, and additional treatments, such as synthetic colloids infusion and FFP administration, were recorded. The presence of SIRS at admission was also evaluated, and the diagnosis was made based on the criteria previously applied in CPV patients , which pâ SPSSâ Statistics Software  was used for the statistical analysis of results. Categorical variables were expressed in absolute and relative frequencies, and for the continuous variables, median and respective range were also measured.The IBMThe association between the clinical variables and outcome (surviving vs. non-surviving) was calculated by the Chi-square test and Fisher’s exact test. For this purpose, continuous variables, such as age, weight, and nadir neutrophil count, were transformed into categorical ones. All variables that showed a significant association (p < 0.05) with the outcome in the univariate analysis were studied using a multivariable logistic regression model to determine whether they could be considered independent prognostic factors.Median, mean, and standard deviation were calculated for the HT. Shapiro–Wilk test of normality was performed and confirmed that HT did not follow the normal distribution. Therefore, Mann–Whitney U and Kruskal–Wallis H tests were carried out to assess their association with the clinical variables. As performed for the outcome, a multivariate analysis was also carried out with the variables that previously showed a statistically significant association with the HT (p < 0.05), using the Cox proportional hazards for this purpose.In this study, 112 dogs were initially identified, but 18 were eventually excluded due to age above the defined limit (n = 1), loss to follow-up (n = 4), uncertainty about the vaccination status (n = 12), and the outcome (n = 1). Therefore, 94 dogs met the final inclusion criteria and were considered for this retrospective study.In our canine population, 52.1% (n = 49) were male and 47.9% (n = 45) were female. The median age was 16 weeks (range: 4–52 weeks) and the median weight at diagnosis was 6.76 kg (range: 0.6–41.4 kg). In terms of dog breed, 48 dogs were purebred and 46 were crossbred. The most frequent purebreds were Labrador (n = 8), Pinscher (n = 5), Estrela Mountain dog (n = 4), German Shepherd (n = 4), and Rafeiro do Alentejo (n = 3), in addition to four breeds with only two dogs each  and to 16 breeds represented by only one patient . For purebred dogs, 18.8% (n = 9) were small sized, 16.7% (n = 8) were medium sized, and 64.6% (n = 31) were large sized, according to the expected adult size and the breed standard.According to prophylactic status, 75.5% (n = 71) had not received any vaccine, 20.2% (n = 19) had been vaccinated, but the initial protocol was still incomplete, and 4.3% (n = 4) had already finished the primo-vaccination schedule at the time of diagnosis.Information about the deworming protocol was available on 84 animals. An adequate deworming schedule was considered when it started at 2 weeks of age and was carried out fortnightly at first and then monthly for year-round control, based on the 2019 AAHA Canine Life Stage Guidelines . Conside9/L with a range from 0.06 to 12.9 × 109/L.Specific clinical signs and blood test results considered of interest for the study were retrieved. Hematochezia was present in most of the dogs included: 71.3% (n = 67). Fever was present in half of the dogs (n = 47) and only 33.0% (n = 31) showed signs related to SIRS at presentation. Serum albumin concentration at admission was available in only 72 patients, 34 of whom had hypoalbuminemia (reference range: 2.7–3.6 g/dL). The median nadir neutrophil count was 1.35 × 10All dogs included were diagnosed with acute viral gastroenteritis, either by a positive fecal antigen test for one of the aforementioned viruses (preferably) or through clinical reasoning, when financial constraints prevented its performance. In the latter cases, only animals that had a strong clinical suspicion toward a viral agent were included, given the clinical presentation, laboratory findings, and response to treatment, in addition to evidence of performing ancillary tests that ruled out other potential causes. It should also be noted that all cases had a clinical diagnosis of CPV enteritis, but as no enzyme-linked immunosorbent assay or polymerase chain reaction test was performed for confirmation in all 94 animals, we chose a more general term, admitting the possibility of a misdiagnosis. In fact, although unlikely, considering the clinical presentation, CDV, enteric coronavirus, or rotavirus may have been responsible for gastroenteritis in some patients, either primarily or in coinfection. In any case, as mentioned, the possibility that the etiology was non-viral was ruled out based on anamnesis and clinical and laboratory findings obtained through physical examination and complementary diagnostic tests .Therefore, the general symptomatic treatment of acute viral gastroenteritis was provided for all patients: Intravenous crystalloid fluid therapy, broad-spectrum antibiotics, antiemetic and gastroprotective medication, and interventional nutrition. In some cases, additional therapies were also administered: 7.4% (n = 7) received synthetic colloids infusion and 26.6% (n = 25) received FFP administration. The median total plasma dose provided was 10 mL/kg with a range from 4.7 to 50 mL/kg. No adverse reactions were described in any of the transfused patients.Patients were kept hospitalized until clinical recovery and were discharged when vomiting and diarrhea had stopped, the appetite had returned to normal and the blood parameters such as packed cell volume, neutrophil count, and albumin normalized.The survival rate was 80.9%: 76 dogs survived and were discharged, but 18 dogs (19.1%) died. Among the patients that died, two were euthanized due to poor prognosis. The median hospitalized time for patients that died was 3½ days (range: 1–8 days). For the surviving patients, the median HT was 5 days (range: 2–16 days).9/L, the presence of fever, and the need for FFP administration were significantly associated with lower survival rate  in the univariate analysis, but only the last two showed a significant association with the outcome in the multivariate analysis (p = 0.021 and p = 0.037) (A nadir neutrophil count lower than 1.35 × 10= 0.037) .The impact of different variables on the HT of dogs that survived was evaluated. One dog of the 76 survivors was excluded because the exact duration of its hospitalization could not be ascertained, although it was confirmed that this patient survived and was discharged from the hospital. Therefore, 75 dogs were evaluated at this stage. In the univariate analysis, HT showed a statistically significant association with the primo-vaccination status, presence of hematochezia, nadir neutrophil count, and plasma administration  . FollowiThe present study retrospectively evaluated a group of young dogs diagnosed with acute presumptive viral gastroenteritis. The main objective was to assess potential prognostic factors associated with HT and survival and to describe this pediatric canine population.The signalment of affected animals was very similar to other recently studied canine populations diagnosed with acute viral gastroenteritis, such as CPV enteritis , 5, 17. The majority of the population was represented by puppies under 6 months of age with an inadequate vaccination protocol. In fact, patients with an inadequate primo-vaccination status needed significantly more time to clinically recover (p = 0.026), probably as a result of a less competent immune response.Hematochezia was one of the most frequent clinical signs observed in affected animals (71.3%). This was an interesting finding because although diarrhea remains one of the most reported signs in patients with viral gastroenteritis, the presence of blood has been described in a lower percentage of patients 21–51%) [1% 5, 1414, 17. Pet al. [et al. [et al. [Fever was present in exactly half of the animals 50%), at the time of presentation, which is a higher relative percentage compared to the previous studies, such as Kalli 0%, at th [et al. , who repOne reason for the differences observed in clinical presentation may be related to the stage of the disease at the time of admission: Possibly, our patients were hospitalized later in the course of the disease, allowing signs such as hematochezia to be already present. Nevertheless, this assumption was not possible to prove because the median duration of presenting complaints before admission could not be calculated for further comparisons. Even so, only 33.0% of our patients showed signs compatible with SIRS, which is in line with the frequency of this condition described by other authors based on the same criteria .et al. [As described in previous studies , 17, hypet al.  in a proet al. , as receIn addition to the supportive treatment provided to all patients, other therapies were described. The FFP was administered in 25 patients, mainly to correct hypoalbuminemia, as previously described . Our stuLimitations of this study include its retrospective nature, the subjective conclusion of viral cause of gastroenteritis, heterogeneity of the patient population, variations in the treatment protocol provided, and incomplete reporting on clinical files. The potential impact of these study weaknesses was minimized by choosing only two referral veterinary hospitals with very similar clinical protocols and work practices and by including only patients with available clinical data and with a clear and known outcome .Another limitation is the diagnostic approach since not all animals performed antigen tests for detection of viruses, such as CPV and CDV. Our diagnosis was mostly clinical, considering the clinical signs, the laboratory abnormalities, the course of the disease, and the treatment response, in addition to the exclusion of the respective differential diagnoses. The treatment of CPV and CDV in young dogs is often expensive for the newly pet owners due to the need of hospitalization for an extended period of time, the aggressive multimodal therapy required, and the regular clinical and laboratory monitoring. For that same reason, an outpatient protocol has already been described for situations where hospitalization cannot be supported by the owner . TherefoThis study illustrates the current state of presumptive viral gastroenteritis and provides new information regarding the impact of various epidemiological, clinical, and therapeutic features on the prognosis. Our main conclusions are: (1) The majority of dogs diagnosed with acute viral gastroenteritis showed an inadequate vaccination status and severe clinical and laboratory abnormalities upon admission; (2) the presence of fever was significantly associated with a lower survival rate; (3) an inadequate primo-vaccination protocol prior admission and hematochezia was significantly associated with longer HT for patients that survived; and (4) the FFP administration in the regimen performed in this subgroup of patients was associated with worst prognosis, making its use in other conditions necessary for a better assessment of its potential therapeutic effect.HG, JA, and FLQ: Conception and design. JA, LR, MJD, and SB: Data collection. TRM, HG, and FLQ: Analysis and interpretation. TRM: Manuscript writing. TRM and FLQ: Reviewed and edited the manuscript. All authors have read and approved the final manuscript.


Spirodela polyrhiza, a species with very low intraspecific sequence variations, including polymorphic NB-ARC-related loci, tubulin-gene-based polymorphism (TBP), simple sequence repeat variations (SSR), multiplexed ISSR genotyping by sequencing (MIG-seq), and low-coverage, reduced-representation genome sequencing (GBS). Four of the five approaches could distinguish 20 to 22 genotypes out of the 23 investigated clones, while TBP resolved just seven genotypes. The choice for a particular method for intraspecific genotyping can depend on the research question and the project budget, while the combination of orthogonal methods may increase the confidence and resolution for the results obtained.The predominantly vegetative propagating duckweeds are of growing commercial interest. Since clonal accessions within a respective species can vary considerably with respect to their physiological as well as biochemical traits, it is critical to be able to track the clones of species of interest after their characterization. Here, we compared the efficacy of five different genotyping methods for  In the last two decades, the evolution of molecular methods has revolutionized phytotaxonomy. These advances have also been applied to the duckweed family, Lemnaceae Martinov ,2. The tPresently, 30 out of 36 species of duckweed can be reliably identified by molecular taxonomy ,8. Thus,Spirodela polyrhiza (L.) Schleid , including four monomorphic clones, were detectable, with all of them originating from the first intron and with no polymorphism scored for the second intron . Since t as well .The SSR method, like the TBP method, uses FLPs for the differentiation of different genotypes. In total, 95 amplification fragments resolved by their mobility on gels were scored as present or absent, which led, again, to no ambiguous characters . The errThe MIG-seq method is an SNP-based method. The final dataset  variations might account for these trait differences, consistent with previous speculations [S. polyrhiza clones from the Southeast Asian ones and clustered clone 9560 from Hungary together with the Asian clones. A great advantage of these two methods is their easy applicability for a wide range of organisms, although both methods require Illumina sequencing and the related equipment [As expected, the methods that yield a high number of total characters and, therefore, differences, i.e., MIG-seq and GBS, showed the best resolution, especially in the phylogeographical studies. The geographical pattern closely fitted the results obtained by whole-genome resequencing in , which aquipment   to about 90 copies [Wolffia australiana for the generation of intraspecific genotyping primer sets.The NB-ARC polymorphism-based genotyping approach has shown a very good performance in genotyping olyrhiza , as obsee genome . To impremaining . In thesemaining  to gener0 copies . This geLemna perpusilla, and tests on a clone of S. intermedia (the sister species of S. polyrhiza) were unsuccessful in the lab of one of the authors (M.B.). The screening of already available whole-genome sequences or shotgun genome information appears to be a straightforward method for developing SSR markers using tools such as GMATA and others . In addition, while the data generated using the four most effective approaches are consistent, to a large extent, and thus provide strong support for their validity, they also suggest the potential advantage of combining two of these orthogonal methods to extend the power of the analysis. For example, using GBS analysis, under a lower stringency of error, only one pair of clones, 9506 and 9316, could not be resolved. These two clones, however, can be distinguished using any of the other three methods. Thus, if data based on GBS and one of these methods can be integrated together in a genotype matrix, the resolution of all 23 clones tested here could become feasible.In summary, we presented five methods, four of which are well-suited for genotyping Spirodela polyrhiza were acquired from the duckweed collection of the Matthias Schleiden Institute—Plant Physiology, University of Jena, Germany, and are available as living materials from the duckweed collections in Jena and at Rutgers State University of New Jersey  under the international four-digit code [rpl16, rps16, and atpF-atpH [3, 0.15 mM KH2PO4, 1 mM MgSO4, 1 mM Ca(NO3)2, 5 μM H3BO3, 0.4 μM Na2MoO4, 13 μM MnCl2, and 25 μM Fe(III)NaEDTA. The plant material was stored at −80 °C for further use.All 23 clones of git code  given ingit code . The clogit code  and genogit code , or thestpF-atpH ,23,36. AtpF-atpH : 8 mM KNDNA was isolated from 100–200 mg fresh weight of duckweed plants using the CTAB method  and quanThe wet lab methods used for the analysis by NB-ARC are described in .TBP amplification was performed by targeting both the 1st and 2nd introns of the multigene β-tubulin family, using two universal primer pairs, Fex-Rex and Fin-Rin, respectively, and PCR conditions described in . FAM-labFor the SSR investigations, a total of 12 SSR markers multiplexed in three sets of four primer pairs each were used, according to . The forFor MIG-seq, 25 µL of each DNA extract at a concentration of 30 ng/µL was sent to LGC Genomics GmbH  for analysis. Sequencing by synthesis was performed with a 150 bp paired-end read chemistry using an Illumina NextSeq500/550 device . Three samples  from the same DNA extracts were run in duplicate for the error estimation. The sequence reads were uploaded to the sequence read archive (SRA) , PRJNA887 cycles, index read 8 cycles) using the Illumina HiSeq 2500 device was performed according to the protocols provided by the manufacturer (Illumina Inc.). Three samples  from the same DNA extracts were run in duplicate for the error estimation. The sequence reads were uploaded to the sequence read archive (SRA) [For the GBS method, DNA was used for the construction of barcoded libraries, essentially performed as described in . Sequencve (SRA) , PRJNA88S. polyrhiza genome available in [The dataset for the analysis by NB-ARC was taken from , and thelable in  was usedWe attempted to make the data analysis for the comparison as similar as possible, although the different methods produced different data types, including FLP (scored as 0/1) and SNP data. Therefore, 0/1 data was recoded as c/a, and each of the five datasets was saved as a FASTA file, which was further processed using R v4.1.0 , with th


Background: The study objective involves the evaluation of preoperative audiovisual information on the operation theater environment as a tool to relieve anxiety in patients posted for infra umbilical elective surgery under spinal anesthesia. Perioperative anxiety is detrimental to both intraoperative hemodynamic instability and postoperative recovery. The design was a single-center, prospective, randomized control trial. There were 120 patients in this study within the 18-60-year-old age range who were American Society of Anaesthesiologists (ASA) class I-II admitted for infra-umbilical surgery excluding lower segment cesarean section (LSCS) under spinal anesthesia. Patients were randomized into two groups: those who were not exposed to an audiovisual tour (NA) (n = 60) and those who were exposed to an audiovisual tour (A) (n = 60). The measurements are based on the demographic details of the patient; the visual analogue score (VAS); and the Amsterdam Preoperative Anxiety and Information Scale (APAIS) for anxiety, hemodynamic parameters i.e., blood pressure (BP), heart rate (HR), respiratory rate (RR), and patient satisfaction score (PSS) on a five-point Likert scale were noted perioperatively for both the groups.Material and methods:Results: The main results showed a significant  decrease in VAS and APAIS for anxiety, HR, and PSS. This was observed during the intraoperative and postoperative periods in the A group in comparison to the NA group.Conclusion: The preoperative audiovisual virtual tour of the operation theater effectively reduces perioperative anxiety and stabilizes HR; it also improves the satisfaction of patients undergoing elective surgery under spinal anesthesia. Anxiety is a feeling of unease, fear, and apprehension in response to any external or internal stimuli that leads to behavioral, emotional, cognitive, or physical symptoms . PreoperThe provision of information to patients plays a significant role in decreasing their anxiety. Patient experiences can be improved by providing them explanations and details of the environment and procedures, highlighting the theater environment. This gives the patient an idea of the function and appearance of the operation theater, making it less unfamiliar and giving the patient an opportunity to ask questions ,8. The tOne of the major goals in the practice of administration of anesthesia is reducing the level of the patient’s anxiety. A preoperative assessment provides this unique opportunity to reduce the patient’s anxiety by building a fine rapport and by providing information regarding the environment of the operation theater, the anesthesia administration, and the surgery procedure. This present study was planned to assess the effects of a preoperative audiovisual virtual tour of the operation theater in decreasing anxiety in patients who are undergoing surgeries with spinal anesthesia.Study designThe present prospective, single-blind randomized control trial was conducted in the Department of Anesthesiology and Critical Care in a tertiary hospital of a teaching institute after obtaining approval from the Biomedical Research Ethics Committee, Pandit Bhagwat Dayal Sharma Post Graduate Institute of Medical Sciences, Rohtak with approval number (EC/NEW/INST/2020/874)(No. BREC/22/013) and CTRI No. CTRI/2022/02/040482 under clinical trial registry, India. Witnessed, informed consent was taken from patients.Patients, endpoints, and assessmentThere were 120 patients in the age group of 18-60 years old who were enrolled in the study; they were American Society of Anaesthesiologists (ASA) class I-II admitted for infra-umbilical (excluding lower segment cesarean section {LSCS}) surgery with spinal anesthesia. Patients who had a prior visit to the operation theater, those who are visual and hearing impaired, those with a history of psychiatric/neurological disorder, those with a history of head injury, those with a history of drug abuse, those with a history of alcohol abuse or any psychological trauma in past six months were excluded from the study. The patients who were scheduled for elective surgery under spinal anesthesia were randomly divided into two groups by permuted block randomization: Group I was composed of patients who were not exposed to the audiovisual tour (NA) (n = 60). Group II was composed of patients exposed to the audiovisual tour (A) (n = 60). Demographic details of the patient (age and sex), blood pressure (BP), heart rate (HR), and respiratory rate (RR) were noted preoperatively. All patients were asked to fill out the six-item questionnaire from APAIS for the assessment of their baseline anxiety: 1. I am worried about the anesthesia, 2. The anesthesia is on my mind continuously, 3. I would like to know as much as possible about the anesthesia, 4. I am worried about the procedure, 5. The procedure is on my mind continuously, 6. I would like to know as much as possible . The meaGroup NA were patients who underwent routine bedside pre-anesthetic consultation the day before the surgery. On the other hand, Group A included the patients who, along with the routine bedside pre-anesthetic consultation, were shown a five minutes video clip containing a virtual tour of the preoperative room, the operation theater, and the recovery room using a tablet with a 10-inch screen size during the pre-anesthetic checkup visit on the day before the surgery. A video clip was accompanied by auditory information related to the operation theater environment like monitors, lights, tables, and staff. A prerecorded video clip was validated and standardized by five anesthesiologists with more than five years of experience in anesthesia administration. On the day of the surgery, APAIS for anxiety was noted before the patients were given anesthesia. The HR, BP, and RR were noted at the start of the anesthesia procedure and repeated every five minutes for 20 minutes and then every 10 minutes until the end of the intraoperative surgery for both of the groups. Postoperatively, 10 mins after shifting from the operation theatre (OT) for both groups, the patients were asked to give an estimate of their anxiety based on the visual analogue score (VAS) and to rate their satisfaction on a scale of 1-5 Likert scale.Statistical analysisAll statistical analyses were performed by using SPSS 22.0 software package . All data were summarized as mean ± SD for continuous variables, numbers, and percentages for categorical variables. The variables were assessed for normality using the Kolmogorov-Smirnov test. A p < 0.05 was accepted as statistically significant.Both the groups were matched for their age; gender; and baseline HR, MAP, RR, VAS, and APAIS for anxiety. The difference observed for these baseline characteristics among all four groups was statistically insignificant Table .VAS and APAIS: The non-audiovisual group and the audiovisual group showed a significant difference in anxiety scores measured with VAS and APAIS as recorded in the preoperative room (timepoint 0) after being shown the prerecorded video of the operation theater with a p-value of 0.001 for both with the mean value of VAS as 30.25 for the NA group and 24.92 for the A group. The mean APAIS at timepoint 0 was 9.65 for group NA and 7.7 for group A  as observed in the preoperative room (time point 0) and at time points 5, 10, 15, 20, 30, and 40 mins post spinal anesthesia. The mean HR for group NA at time points 0, 5, 10, 15, 20, 30, and 40 mins were 87.48, 86.55, 85.13, 84.10, 82.88, 81.47, and 80.17, respectively. The mean HR for group A at time points 0, 5, 10, 15, 20, 30, and 40 mins were 79.62, 79.20, 78.50, 78.08, 77.57, 76.93, and 76.45, respectively  except for the stabilization of HR. Other factors like better interaction through audiovisual activity and psychological support could have contributed to the reduction in anxiety, along with their familiarization with the environment. More studies with large sample sizes and more specific patient populations are required to exactly establish the relationship between environment familiarization and perioperative anxiety.The limitations of our study include being single-centric, having a small sample size, the nonuniformity in nature of surgeries, and the nonassessment of the subjective anxiety scores intraoperatively. More elaborate subjective scores for anxiety and biochemical markers  can be used to better correlate variations in anxiety levels.From the observations gathered, it is evident that showing patients a prerecorded audiovisual clip of the operation theater effectively reduces perioperative anxiety, stabilizes HR, and improves the satisfaction score of patients who undergo elective surgery under regional anesthesia. This method is simple, easy to practice, has low cost, and can increase overall subjective patient satisfaction, potentially improving patient safety and clinical outcomes.