Analysis of relevant factors affecting the prognosis of mechanical thrombectomy in patients with acute posterior circulation large vessel occlusion cerebral infarction.

OBJECTIVE: To explore the factors affecting the prognosis of patients with acute posterior circulation large vessel occlusion cerebral infarction (PCO) after mechanical thrombectomy. METHOD: A retrospective study was conducted on a total of 58 patients who received thrombectomy and presented within 24 h of onset with PCO from 31 September 2020 to 31 December 2022. They were divided into two groups based on a 90-day mRS score(The mRS score of 0-3 was defined as a good prognosis, and 4-6 was defined as a poor prognosis).A univariate analysis was conducted on baseline data such as age and patient past medical history, as well as extended cerebral infarction thrombolysis grade (eTICI grade) and incidence of symptomatic intracranial hemorrhage (sICH) after surgery, for the groups with good prognosis and poor prognosis. Factors affecting the 90-day prognosis of patients were also analyzed in subgroups. RESULTS: The preoperative National Institutes of Health Stroke Scale (NIHSS score)[21(12-35) vs 35(35-35)], postoperative 24-h NIHSS score[13(8-22) vs 35(35-35)], computed tomography (CT)[9(9-10) vs 6.5(6-7.75)] and computed tomography (CTP) brain blood volume (CBV)[9(8-10) vs 4(2-7.75)], cerebral blood flow (CBF)[7(4.5-9) vs 2(1-4)], time to peak (Tmax) [1(0.5-4) vs 0(0-1.75)] imaging of the posterior circulation Alberta stroke project early CT score (pc-ASPECTS score), Different locations of vascular occlusion, time from femoral artery puncture to vascular recanalization(64.96 ± 33.47 vs 92.68 ± 53.17). The differences in the conversion rate of postoperative intracranial hemorrhage(0 vs 16.1%) and the incidence of sICH(0 vs 12.9%) were statistically significant (P < 0.05). The subgroup analysis showed that vascular occlusion site, preoperative CBV pc-ASPECTS scores, and postoperative sICH occurrence were related to the 90-day prognosis of patients, and the differences were statistically significant (P < 0.05). CONCLUSIONS: Some factors that can affect the prognosis of mechanical thrombectomy in patients with acute posterior circulation large vessel occlusion cerebral infarction. Preoperative clinical symptoms and imaging evaluation have certain evaluation values for prognosis.

Single and mixed associations of composite antioxidant diet on triglyceride-glucose index.

BACKGROUND: Although the relationship between oxidative stress and insulin resistance (IR) has been established, the associations of the composite dietary antioxidant index (CDAI) and its components with the surrogate index of insulin resistance (IR), triglyceride-glucose index (TyG), is still not clear. METHODS: This study analyzed the cross-sectional data of the National Health and Nutrition Examination Survey (NHANES) from 2003 to 2018. Multivariate linear regression, weighted quantile sum (WQS) regression, and Bayesian kernel machine regression (BKMR) were used to analyze the associations of the CDAI and its components with the TyG. In addition, subgroup analysis and several sensitivity analyses were conducted. RESULTS: A total of 14,673 participants with complete data were included, with a median age of 50 years and 7,257 women (49%). Multivariate linear regression showed that after full adjustment, the CDAI was significantly negatively associated with the TyG [ß: -0.005, 95% CI: (-0.008, -0.002), p = 0.002]. The model in which six nutrients were mutually corrected showed that vitamin E (per-SD increase) was most strongly associated with the TyG [ß: -0.062, 95% CI: (-0.074, -0.050), p < 0.0001]. In the WQS model, the WQS index of the antioxidant diet was negatively associated with the TyG (ß: -0.060; P < 0.0001). Similar effects were observed in the BKMR analysis. Notably, in the WQS and BKMR models, vitamin E became the most influential component. In addition, in the subgroup analysis, the association between the CDAI and the TyG in overweight or obese and diabetic populations was significantly weaker. CONCLUSION: Antioxidant diets, especially vitamin E, are significantly negatively correlated with TyG. This study emphasizes the important value of supplementing vitamin E to improve IR. However, patients with poor weight management and diabetes seem to benefit less from antioxidant diets.

The value of the platelet/high-density lipoprotein cholesterol ratio in predicting depression and its cardiovascular disease mortality: a population-based observational study.

Background: Cardiovascular disease (CVD) and depression have a bidirectional association, with inflammation and metabolic factors being common important triggers for both conditions. However, as a novel inflammatory and metabolic marker, platelet-to-HDL-C ratio (PHR) has not been established in relation to depression and cardiovascular disease. Materials and methods: Participants aged 20 years and older were included in the 2005-2018 NHANES database. PHR was calculated as the ratio of platelet count (1000 cells/µL) to HDL-C (mmol/L). The Patient Health Questionnaire (PHQ-9) was used to diagnose depression, with a cutoff value of 10. Weighted logistic regression analysis and restricted cubic spline (RCS) analysis were employed to examine the association between PHR and depression-related features. Additionally, weighted COX regression and RCS were used to analyze the association of PHR with CVD mortality in patients with depression. Receiver operating characteristic curves were used to assess whether PHR had an advantage over HDL-C in predicting depression. Finally, the mediating role of PHR in the latest cardiovascular health indicator Life's Essential 8 and depression was explored. Results: A total of 26,970 eligible participants were included, including 2,308 individuals with depression, representing approximately 160 million U.S. adults when weighted. After full adjustment, we estimated that the odds ratio (OR) of depression associated with a per standard deviation (SD) increase in PHR was 1.06 (95% CI: 1.01-1.12, P=0.03). The restricted cubic spline (RCS) analysis indicated a linear association (Nonlinear P=0.113). When PHR was divided into four groups based on quartiles and included in the model after full adjustment for depression risk factors, participants in quartile 2, quartile 3, and quartile 4 of PHR showed a trend of increasing risk of depression compared to the lowest quartile group (P trend=0.01). In addition, weighted COX regression and RCS revealed that a per SD increase in PHR was associated with a higher risk of CVD mortality among patients with depression (HR: 1.38, 95% CI: 1.05-1.81, P=0.02, Nonlinear P=0.400). Subgroup analyses showed that current alcohol consumption enhanced the association between PHR and depression (P for interaction=0.017). Furthermore, the areas under the ROC curves (AUC) were 0.556 (95% CI, 0.544-0.568; P < 0.001) for PHR and 0.536 (95% CI, 0.524-0.549; P < 0.001) for HDL-C (PDeLong = 0.025). Finally, mediation analysis indicated that PHR was an intermediate mechanism between LE8 and depression (mediation proportion=5.02%, P=0.02). Conclusion: In U.S. adults, an increase in PHR linearly increases the risk of depression and CVD mortality among individuals with depression. Additionally, PHR has a better predictive advantage for depression compared to HDL-C. Furthermore, PHR significantly mediates the association between LE8 scores and depression.

Type II diabetes mellitus increases the risk of inflammatory bowel disease in a prospective cohort study.

BACKGROUND: Inflammatory bowel disease is a common digestive disorder and diabetes can lead to intestinal dysfunction. Patients with inflammatory bowel disease in combination with diabetes present a higher rate of hospitalization and consumption of medical resources, yet the association between type 2 diabetes and Inflammatory bowel disease remains unknown. METHODS: We studied 313,008 participants from the UK Biobank, including 5891 patients with type 2 diabetes at baseline. Multivariate Cox proportional risk models were constructed to examine the risks associated with type 2 diabetes and inflammatory bowel disease and its subtypes (Crohn's disease, ulcerative colitis). Potential confounders including sociodemographic, lifestyle, physical body indicators, psychological state, hypertension, and thyroid-related disorders were adjusted. Propensity score matching was also performed to analyze their sensitivity. RESULTS: Of a total of 313,008 participants included in the study, 5891 (1.88 %) were diagnosed with type 2 diabetes mellitus at baseline and 1829 (0.58 %) of the entire cohort developed inflammatory bowel disease during follow-up, with a median follow-up time of 13.72 years. Patients with type 2 diabetes had a higher cumulative risk of inflammatory bowel disease compared to the non-type 2 diabetes population (inflammatory bowel disease: 1.24% vs. 0.57%, p < 0.001; Crohn's disease: 0.46% vs. 0.15%, p < 0.001; ulcerative colitis: 0.73% vs. 0.35%, p < 0.001). Multivariate Cox regression analysis showed that type 2 diabetes was independently associated with inflammatory bowel disease (Hazard Ratio: 1.61 [95% Confidence Interval: 1.26-2.06], p < 0.001), Crohn's disease (Hazard Ratio: 2.10 [95% Confidence Interval: 1.39-3.17], p < 0.001) and ulcerative colitis (Hazard Ratio: 1.58 [95% Confidence Interval: 1.15-2.18], p = 0.005). In a propensity-matched analysis, type 2 diabetes still showed its ability to predict the risk of inflammatory bowel disease (Hazard Ratio: 2.09 [95% Confidence Interval: 1.55-2.83], p < 0.001), Crohn's disease (Hazard Ratio: 3.49 [95% Confidence Interval: 2.00 to 6.09], p < 0.001), and ulcerative colitis (Hazard Ratio: 1.76 [95% Confidence Interval: 1.20 to 2.56], p = 0.003) of robustness. CONCLUSION: In patients with type 2 diabetes mellitus, the risk of inflammatory bowel disease is higher, and the presence of gastrointestinal symptoms in patients with type 2 diabetes requires vigilance for the possibility of inflammatory bowel disease in clinical practice.

The association of the platelet/high-density lipoprotein cholesterol ratio with self-reported stroke and cardiovascular mortality: a population-based observational study.

BACKGROUND: Previous studies have shown that the relationship between high-density lipoprotein cholesterol (HDL-C) and stroke is controversial, and the association between the platelet/high-density lipoprotein cholesterol ratio (PHR), a novel marker for inflammation and hypercoagulability states, and stroke has not been established. METHODS: This study presents an analysis of cross-sectional data from the 2005-2018 National Health and Nutrition Examination Survey (NHANES). Stroke history, HDL-C levels, and platelet counts were obtained during cross-sectional surveys. The PHR was calculated as the ratio of the number of platelets to HDL-C concentration. Weighted logistic regression was used to assess the associations of HDL-C and the PHR with stroke. Nonlinearity of this relationship was determined through restricted cubic splines (RCSs) and two-piecewise linear regression for identifying inflection points. Furthermore, Cox regression was utilized to prospectively analyze the associations of the PHR and HDL-C concentration with cardiovascular disease (CVD) mortality in stroke survivors. RESULTS: A total of 27,301 eligible participants were included in the study; mean age, 47.28 years and 50.57% were female, among whom 1,040 had a history of stroke. After full adjustment, the odds ratio (OR) of stroke associated with a per standard deviation (SD) increase in the PHR was estimated at 1.13 (95% confidence interval (CI): 1.03 - 1.24, P = 0.01), and the OR of stroke associated with a per SD increase in HDL-C was 0.95 (95% CI: 0.86-1.05, P = 0.30). The RCS indicated a nonlinear relationship for both variables (PPHR = 0.018 and PHDL-C = 0.003), and further piecewise linear regression identified inflection points at PHR = 223.684 and HDL-C = 1.4 mmol/L. Segmental regression indicated that in the PHR ≥ 223.684 segment, the estimated OR of stroke associated with a per-SD increase in the PHR was 1.20 (95% CI: 1.09 - 1.31, P < 0.001), while the association of stroke with HDL-C was not significant before or after the inflection point (P > 0.05). Furthermore, Cox regression and RCS showed that a per-SD increase in the PHR was linearly associated with a greater risk of CVD mortality among stroke survivors (HR: 1.14, 95% CI: 1.06 - 1.22, P < 0.001; nonlinear, P = 0.956), while HDL-C was not significantly associated with CVD mortality. CONCLUSION: The association between the PHR and stroke incidence exhibited a significant threshold effect, with an inflection point at 223.684. A PHR exceeding 223.684 was positively associated with stroke, while the association between HDL-C and stroke was not significant. Additionally, the PHR was positively and linearly associated with CVD mortality among stroke survivors.

Towards a hybrid model-driven platform based on flux balance analysis and a machine learning pipeline for biosystem design.

Metabolic modeling and machine learning (ML) are crucial components of the evolving next-generation tools in systems and synthetic biology, aiming to unravel the intricate relationship between genotype, phenotype, and the environment. Nonetheless, the comprehensive exploration of integrating these two frameworks, and fully harnessing the potential of fluxomic data, remains an unexplored territory. In this study, we present, rigorously evaluate, and compare ML-based techniques for data integration. The hybrid model revealed that the overexpression of six target genes and the knockout of seven target genes contribute to enhanced ethanol production. Specifically, we investigated the influence of succinate dehydrogenase (SDH) on ethanol biosynthesis in Saccharomyces cerevisiae through shake flask experiments. The findings indicate a noticeable increase in ethanol yield, ranging from 6 % to 10 %, in SDH subunit gene knockout strains compared to the wild-type strain. Moreover, in pursuit of a high-yielding strain for ethanol production, dual-gene deletion experiments were conducted targeting glycerol-3-phosphate dehydrogenase (GPD) and SDH. The results unequivocally demonstrate significant enhancements in ethanol production for the engineered strains Δsdh4Δgpd1, Δsdh5Δgpd1, Δsdh6Δgpd1, Δsdh4Δgpd2, Δsdh5Δgpd2, and Δsdh6Δgpd2, with improvements of 21.6 %, 27.9 %, and 22.7 %, respectively. Overall, the results highlighted that integrating mechanistic flux features substantially improves the prediction of gene knockout strains not accounted for in metabolic reconstructions. In addition, the finding in this study delivers valuable tools for comprehending and manipulating intricate phenotypes, thereby enhancing prediction accuracy and facilitating deeper insights into mechanistic aspects within the field of synthetic biology.

Mucus-Penetrating Nanoassembly as Potential Oral Phototherapeutic Formulation against Multi-Drug Resistant Helicobacter pylori Infection.

Helicobacter pylori (H. pylori) infection presents increasing challenges to antibiotic therapies in limited penetration through gastric mucus, multi-drug resistance (MDR), biofilm formation, and intestinal microflora dysbiosis. To address these problems, herein, a mucus-penetrating phototherapeutic nanomedicine (RLs@T780TG) against MDR H. pylori infection is engineered. The RLs@T780TG is assembled with a near-infrared photosensitizer T780T-Gu and an anionic component rhamnolipids (RLs) for deep mucus penetration and light-induced anti-H. pylori performances. With optimized suitable size, hydrophilicity and weak negative surface, the RLs@T780TG can effectively penetrate through the gastric mucus layer and target the inflammatory site. Subsequently, under irradiation, the structure of RLs@T780TG is disrupted and facilitates the T780T-Gu releasing to target the H. pylori surface and ablate multi-drug resistant (MDR) H. pylori. In vivo, RLs@T780TG phototherapy exhibits impressive eradication against H. pylori. The gastric lesions are significantly alleviated and intestinal bacteria balance is less affected than antibiotic treatment. Summarily, this work provides a potential nanomedicine design to facilitate in vivo phototherapy in treatment of H. pylori infection.

Circulating IL-17 reduces the risk of cisplatin-induced hearing loss in children: a bidirectional two-sample Mendelian randomization study.

Studies have reported that cytokines and their related signaling pathways play a role in inner ear diseases. In clinical practice, approximately 50% of pediatric cancer patients experience irreversible hearing loss after cisplatin treatment. However, currently, there is a lack of systematic research on the causal relationship between circulating cytokines and cisplatin-induced hearing loss in children. Genetic variant data for 41 circulating cytokines were obtained from a meta-analysis of genome-wide association studies (GWAS) among 8293 individuals of Finnish descent. The GWAS data for Cisplatin-induced hearing loss in children were derived from a multicenter cohort of European pediatric cancer patients and survivors (N = 390), including both cases with hearing loss after cisplatin chemotherapy and controls without hearing loss. Multiple methods were employed for bidirectional Mendelian randomization (MR) estimation. Bonferroni correction was applied to adjust the original P-values, followed by a series of sensitivity analyses. In the directional Mendelian randomization (MR) analysis, it was found that IL-17 was significantly associated with a reduced risk of Cisplatin-induced hearing loss in children (OR: 0.18, CI: 0.06-0.48, P < 0.001, FDR = 0.041). In the reverse MR analysis, there were some nominal causal relationships of Cisplatin-induced hearing loss in children with certain cytokines [M-CSF: (OR: 1.04, CI: 1.01-1.08, P = 0.010, FDR = 0.41); IL-2RA: (OR: 1.03, CI: 1.00-1.05, P = 0.044, FDR = 0.447); MIP-1ß: (OR: 1.02, CI: 1.00-1.04, P = 0.041, FDR = 0.447)]. leave-one-out analysis demonstrated that only M-CSF exhibited stability. These findings reveal a causal relationship between IL-17 and cisplatin-induced hearing loss in children. Further research is needed to determine the potential protective mechanisms of IL-17 in cisplatin-induced ototoxicity.

Predictive value of fibrinogen levels for 90-day functional outcomes after intravenous thrombolysis in patients with acute ischaemic stroke.

PURPOSE: We aimed to investigate the correlation between fibrinogen levels and functional outcomes at 90 days after intravenous thrombolysis therapy (IVT) in patients with acute ischaemic stroke (AIS). METHODS: We identified patients with AIS who received IVT (alteplase 0.6 or 0.9 mg/kg) between 1 January 2019 and 31 March 2022 in Yancheng 1st People's Hospital. Fibrinogen levels were measured before IVT, and the 90-day post-stroke functional outcome was evaluated using the modified Rankin Scale (mRS). An mRS score of 0-2 indicated functional independence, whereas an mRS score of 3-6 indicated functional dependence. Potential outcome predictors were evaluated using univariate and multivariate analyses, and receiver operating characteristic (ROC) curve analysis was performed to assess the performance of fibrinogen levels in predicting the 90-day outcome. RESULTS: A total of 276 patients with AIS who received IVT within 4.5 h of stroke onset were enrolled, of whom 165 and 111 were categorised into the functional independence and functional dependence groups, respectively. Univariate analysis showed that the fibrinogen, homocysteine, high-density lipoprotein cholesterol, and D-dimer levels; age; National Institutes of Health Stroke Scale (NIHSS) score on admission; NIHSS score 24 h after IVT; and incidence of cardioembolism were higher in the functional dependence group than in the functional independence group (P < 0.05). Meanwhile, the thrombin time and the incidence of small-vessel occlusion in the functional dependence group were smaller than those in the functional independence group (P < 0.05). Multivariate logistic regression analysis showed that fibrinogen and homocysteine levels were both independent risk factors for 90-day functional dependence in patients with AIS (odds ratio [OR] 2.822, 95% confidence interval [95% CI]: 1.214-6.558, P = 0.016 for fibrinogen; OR 1.048, 95 %CI: 1.002-1.096, P = 0.041 for homocysteine). The area under the ROC curve of fibrinogen levels before IVT for predicting poor functional outcomes was 0.664, with a sensitivity, specificity, positive predictive value, and negative predictive value of 40.9%, 80.8%, 68.9%, and 64.3%, respectively. CONCLUSION: In patients with AIS, fibrinogen levels have a certain predictive value for short-term functional outcomes after IVT.

Alleviating glucose repression and enhancing respiratory capacity to increase itaconic acid production.

The Crabtree effect products ethanol and acetic acid can be used for itaconic acid (IA) production in Saccharomyces cerevisiae. However, both the IA synthesis and oxidative phosphorylation pathways were hampered by glucose repression when glucose was used as the substrate. This study aimed to improve IA titer by increasing gene expressions related to glucose derepression without impairing yeast growth on glucose. Engineering the acetyl-CoA synthesis pathway increased the titer of IA to 257 mg/L in a urea-based medium. Instead of entire pathway overexpression, we found that some signaling pathways regulating glucose repression were effective targets to improve IA production and respiratory capacity. As a consequence of the reduced inhibition, IA titer was further increased by knocking out a negative regulator of the mitochondrial retrograde signaling MKS1. SNF1/MIG1 signaling was disturbed by deleting the hexokinase HXK2 or an endoplasmic reticulum membrane protein GSF2. The shaking results showed that XYY286 (BY4741, HO::cadA, Y::Dz.ada, 208a::Mt.acs, Δhxk2, pRS415-cadA, pRS423-aac2) accumulated 535 mg/L IA in 168 h in the YSCGLU medium. qRT-PCR results verified that deletion of MKS1 or HXK2 upregulated the gene expressions of the IA synthesis and respiratory pathways during the growth on glucose.

Utilization of ethanol for itaconic acid biosynthesis by engineered Saccharomyces cerevisiae.

In Saccharomyces cerevisiae, ethanol can serve as both a carbon source and NADH donor for the production of acetyl-CoA derivatives. Here we investigated the metabolic regulation of ethanol utilization for itaconic acid production by S. cerevisiae. To understand the interconnection between the TCA cycle and the glyoxylate pathway, mitochondrial membrane transporter proteins SFC1, YHM2, CTP1, DIC1 and MPC1 were knocked out and results showed that SFC1 functions as an important entrance of the glyoxylate pathway into the TCA cycle, and YHM2 is helpful to IA production but not the primary pathway for citric acid supply. To decrease the accumulation of acetic acid, the major ADP/ATP carrier of the mitochondrial inner membrane, AAC2, was upregulated and determined to accelerate ethanol utilization and itaconic acid production. RNA sequencing results showed that AAC2 overexpression enhanced IA titer by upregulating the ethanol-acetyl-CoA pathway and NADH oxidase in the mitochondrial membrane. RNA-seq analysis also suggested that aconitase ACO1 may be a rate-limiting step of IA production. However, the expression of exogenous aconitase didn't increase IA production but enhanced the rate of ethanol utilization and decreased cell growth.

A rare case of mesenteric heterotopic pancreas presenting as an inflammatory mass.

Heterotopic pancreas (HP) is a rare condition, is commonly asymptomatic and found incidentally at autopsy or during abdominal surgery. HP is usually found in the upper gastrointestinal tract, particularly in the stomach, duodenum and jejunum. Mesenteric heterotopic pancreas (MHP) has been rarely described in the literature, and there has been no reported case under ten years of age. We report a unique case of a 7-year-old African boy who presented with clinical appendicitis and intraoperatively was found to have a necrotic nodular mass in the jejunal mesentery which required segmental small bowel resection. Histology revealed mesenteric heterotopic pancreas with normal pancreatic tissue. As pre-operative diagnosis of MHP is difficult; we encourage a heightened awareness of this condition as a differential diagnosis in children presenting with abdominal pain and an inflammatory mass, which will subsequently aid in the management of these patients.

A rare case of multi-focal non-contiguous necrotizing soft tissue infections and literature review.

Necrotising soft tissue infection (NSTI) is a rare yet life-threatening surgical emergency, characterized by rapidly spreading infection below the epidermis to the soft tissue layers of the dermis, subcutaneous layers, fascia and sometimes muscle. Multi-focal NSTIs have been rarely described in the literature, with reported cases predominantly involving immunocompromised patients. We report a case of an immunocompromised 70-year-old woman who initially presented with concern of a perforated colonic malignancy requiring exploratory laparotomy and within 24 hours while on inotropes, developed rapidly progressing necrotizing fasciitis (NF) of the left thigh with renal failure secondary to sepsis. Despite aggressive debridement, a progress computed tomography later showed multi-focal non-contiguous necrotising myositis of the whole left lower limb and right gluteal regions. Early diagnosis of multi-focal NF especially in immunocompromised patients, repeat assessment, aggressive surgical debridement and prompt antibiotics usage are the key to treatment.

A rare presentation of an ectopic kidney with pyelonephritis mimicking appendicitis.

Acute appendicitis is one of the top differential diagnoses of right lower quadrant pain in the emergency department. There are many other conditions that may mimic appendicitis such as diverticulitis, colitis and gynecological conditions. We report a rare diagnosis of a patient who presents with characteristic clinical and laboratory features of appendicitis with severe sepsis, but later showed acute pyelonephritis of a malrotated right ectopic kidney on computer tomography. An ectopic kidney is very rare with an incidence of 1 in 3000. It is usually asymptomatic, although it may also associate with obstruction, infection and urolithiasis. This case report raises the importance of early recognition of the correct diagnosis using imaging in appropriate clinical settings, and prompt antibiotic treatment can avoid unnecessary surgical intervention, preserve renal function and prevent a life-threatening catastrophe.

Integration of Metabolite Profiling and Transcriptome Analysis Reveals Genes Related to Volatile Terpenoid Metabolism in Finger Citron (C. medica var. sarcodactylis).

Finger citron (Citrus medica var. sarcodactylis) is a popular ornamental tree and an important source of essential oils rich in terpenoids, but the mechanisms behind volatile formation are poorly understood. We investigated gene expression changes combined with volatile profiling of ten samples from three developing organs: flower, leaf, and fruit. A total of 62 volatiles were identified with limonene and γ-terpinene being the most abundant ones. Six volatiles were identified using partial least squares discriminant analysis (PLS-DA) that could be used as markers for distinguishing finger citron from other citrus species. RNA-Seq revealed 1,611,966,118 high quality clean reads that were assembled into 32,579 unigenes. From these a total of 58 terpene synthase (TPS) gene family members were identified and the spatial and temporal distribution of their transcripts was measured in developing organs. Transcript levels of transcription factor genes AP2/ERF (251), bHLH (169), bZIP (76), MYB (155), NAC (184), and WRKY (66) during finger citron development were also analyzed. From extracted subnetworks of three modules constructed by weighted gene co-expression network analysis (WGCNA), thirteen TPS genes and fifteen transcription factors were suggested to be related to volatile terpenoid formation. These results provide a framework for future investigations into the identification and regulatory network of terpenoids in finger citron.

Intra-abdominal tuberculosis masquerading as ovarian carcinoma.

Intra-abdominal tuberculosis (TB) is rare in Australia, but it can be found in those who emigrate from endemic areas. We report a rare case of a 28-year-old lady from the Philippines who presented with abdominal pain, ascites and an elevated cancer antigen (CA) 125 with an initial concern of ovarian malignancy. She underwent a diagnostic laparoscopy which revealed typical features of intra-abdominal TB and histological features of granulomatous inflammation. The symptoms and signs of intra-abdominal TB are non-specific and can mimic many other conditions. The gold-standard mode of diagnosis in intra-abdominal TB is laparoscopy with tissue biopsy. Ovarian malignancy is relatively rare in pre-menopausal women; hence an elevated CA 125 warrants a broader differential diagnosis. It is important to have intra-abdominal TB as a differential even in the non-endemic settings to avoid delay in diagnosis and appropriate management.

Transcription factor CitERF71 activates the terpene synthase gene CitTPS16 involved in the synthesis of E-geraniol in sweet orange fruit.

The unique flavor of Citrus fruit depends on complex combinations of soluble sugars, organic acids, and volatile compounds. The monoterpene E-geraniol is an important volatile, contributing to flavor in sweet orange (Citrus sinensis Osbeck). Moreover, antifungal activity of E-geraniol has also been observed. However, the terpene synthase (TPS) responsible for its synthesis has not been identified in sweet orange. Terpene synthase 16 (CitTPS16) was shown to catalyze synthesis of E-geraniol in vitro, and transient overexpression of CitTPS16 in fruits and leaves of Newhall sweet orange resulted in E-geraniol accumulation in vivo. Having identified the responsible enzyme, we next examined transcriptional regulation of CitTPS16 in the fruit. Among cloned members of the AP2/ERF transcription factor gene family, CitERF71 showed a similar expression pattern to CitTPS16. Moreover, CitERF71 was able to activate the CitTPS16 promoter based on results from transient dual-luciferase assays and yeast one-hybrid assays. EMSAs showed that CitERF71 directly binds to ACCCGCC and GGCGGG motifs in the CitTPS16 promoter. These results indicate an important role for CitERF71 in transcriptional regulation of CitTP16 and, therefore, in controlling production of E-geraniol in Citrus fruit.

Genome-Wide Identification, Expression Patterns, and Functional Analysis of UDP Glycosyltransferase Family in Peach (Prunus persica L. Batsch).

Peach (Prunus persica L. Batsch) is a commercial grown fruit trees, important because of its essential nutrients and flavor promoting secondary metabolites. The glycosylation processes mediated by UDP-glycosyltransferases (UGTs) play an important role in regulating secondary metabolites availability. Identification and characterization of peach UGTs is therefore a research priority. A total of 168 peach UGT genes that distributed unevenly across chromosomes were identified based on their conserved PSPG motifs. Phylogenetic analysis of these genes with plant UGTs clustered them into 16 groups (A-P). Comparison of the patterns of intron-extron and their positions within genes revealed one highly conserved intron insertion event in peach UGTs. Tissue specificity, temporal expression patterns in peach fruit during development and ripening, and in response to abiotic stress UV-B irradiation was investigated using RNA-seq strategy. The relationship between UGTs transcript levels and concentrations of glycosylated volatiles was examined to select candidates for functional analysis. Heterologous expressing these candidate genes in Escherichia coli identified UGTs that were involved in the in vitro volatile glycosylation. Our results provide an important source for the identification of functional UGT genes to potential manipulate secondary biosynthesis in peach.

Comparative analysis of flower volatiles from nine citrus at three blooming stages.

Volatiles from flowers at three blooming stages of nine citrus cultivars were analyzed by headspace-solid phase microextraction (HS-SPME)-GC-MS. Up to 110 volatiles were detected, with 42 tentatively identified from citrus flowers for the first time. Highest amounts of volatiles were present in fully opened flowers of most citrus, except for pomelos. All cultivars were characterized by a high percentage of either oxygenated monoterpenes or monoterpene hydrocarbons, and the presence of a high percentage of nitrogen containing compounds was also observed. Flower volatiles varied qualitatively and quantitatively among citrus types during blooming. Limonene was the most abundant flower volatile only in citrons; α-citral and ß-citral ranked 2nd and 3rd only for Bergamot, and unopened flowers of Ponkan had a higher amount of linalool and ß-pinene while much lower amount of γ-terpinene and p-cymene than Satsuma. Taking the average of all cultivars, linalool and limonene were the top two volatiles for all blooming stages; ß-pinene ranked 3rd in unopened flowers, while indole ranked 3rd for half opened and fully opened flower volatiles. As flowers bloomed, methyl anthranilate increased while 2-hexenal and p-cymene decreased. In some cases, a volatile could be high in both unopened and fully opened flowers but low in half opened ones. Through multivariate analysis, the nine citrus cultivars were clustered into three groups, consistent with the three true citrus types. Furthermore, an influence of blooming stages on clustering was observed, especially with hybrids Satsuma and Huyou. Altogether, it was suggested that flower volatiles can be suitable markers for revealing the genetic relationships between citrus cultivars but the same blooming stage needs to be strictly controlled.