Plant-Based Meat Analogs and Their Effects on Cardiometabolic Health: An 8-Week Randomized Controlled Trial Comparing Plant-Based Meat Analogs With Their Corresponding Animal-Based Foods.

BACKGROUND: With the growing popularity of plant-based meat analogs (PBMAs), an investigation of their effects on health is warranted in an Asian population. OBJECTIVES: This research investigated the impact of consuming an omnivorous animal-based meat diet (ABMD) compared with a PBMAs diet (PBMD) on cardiometabolic health among adults with elevated risk of diabetes in Singapore. METHODS: In an 8-wk parallel design randomized controlled trial, participants (n = 89) were instructed to substitute habitual protein-rich foods with fixed quantities of either PBMAs (n = 44) or their corresponding animal-based meats (n = 45; 2.5 servings/d), maintaining intake of other dietary components. Low-density lipoprotein (LDL) cholesterol served as primary outcome, whereas secondary outcomes included other cardiometabolic disease-related risk factors (e.g. glucose and fructosamine), dietary data, and within a subpopulation, ambulatory blood pressure measurements (n = 40) at baseline and postintervention, as well as a 14-d continuous glucose monitor (glucose homeostasis-related outcomes; n = 37). RESULTS: Data from 82 participants (ABMD: 42 and PBMD: 40) were examined. Using linear mixed-effects model, there were significant interaction (time × treatment) effects for dietary trans-fat (increased in ABMD), dietary fiber, sodium, and potassium (all increased in PBMD; P-interaction <0.001). There were no significant effects on the lipid-lipoprotein profile, including LDL cholesterol. Diastolic blood pressure (DBP) was lower in the PBMD group (P-interaction=0.041), although the nocturnal DBP dip markedly increased in ABMD (+3.2% mean) and was reduced in PBMD (-2.6%; P-interaction=0.017). Fructosamine (P time=0.035) and homeostatic model assessment for ß-cell function were improved at week 8 (P time=0.006) in both groups. Glycemic homeostasis was better regulated in the ABMD than PBMD groups as evidenced by interstitial glucose time in range (ABMD median: 94.1% (Q1:87.2%, Q3:96.7%); PBMD: 86.5% (81.7%, 89.4%); P = 0.041). The intervention had no significant effect on the other outcomes examined. CONCLUSIONS: An 8-wk PBMA diet did not show widespread cardiometabolic health benefits compared with a corresponding meat based diet. Nutritional quality is a key factor to be considered for next generation PBMAs. This trial was registered at https://clinicaltrials.gov/as NCT05446753.

Addressing the functional needs of left ventricular assist device candidates: Development and feasibility of an occupational therapy pre-operative evaluation.

BACKGROUND: Everyday living with a left ventricular assist device (LVAD) is complex, particularly for people with physical or cognitive impairments or limited social supports. There is a need for standardized pre-operative functional evaluations. OBJECTIVES: Our objectives were to describe a pre-operative occupational therapy (OT) evaluation for LVAD candidates, assess its feasibility in routine care, and characterize functional needs. METHODS: We retrospectively reviewed electronic medical records of pre-operative OT consultations for LVAD candidates over four years (n = 209). Occupational profile, vision, and sensation were operationalized from documentation narratives. Daily functioning was measured with Activity Measure for Post-Acute Care, grip strength with dynamometer, cognition with Montreal Cognitive Assessment and Allen Cognitive Level Screen-5, and LVAD self-management with a performance-based ordinal scale. RESULTS: 89.5 % of consultations were completed, averaging 61.2 min (n = 187): 79.1 % (148/187) inpatient and 20.9 % (39/187) outpatient. Patients completed 87.7 % (164/187) to 100.0 % (187/187) of evaluation components. 21.9 % (41/187) of candidates lived alone. 6.4 % (12/187) and 7.0 % (13/185) had visual and sensory dysfunction. 57.4 % were independent with daily activities (104/181). 17.7 % (32/181) had impaired grip strength. 69.3 % (124/179) had impaired cognition, 29.7 % (51/172) with impaired functional cognition for everyday activities. 88.4 % (145/164) required physical or cueing assistance while practicing LVAD batteries management. OTs interpreted that 20.9 % (39/187) would likely require 24/7 post-operative support with LVAD self-care. CONCLUSION: Pre-operative OT evaluations were feasible and emphasized complex functional needs. Assessing LVAD self-care abilities may inform candidacy and facilitate early interventions to optimize functioning. OT should be consulted within interprofessional teams for all LVAD candidates.

Cognitive Demands Influence Upper Extremity Motor Performance During Recovery From Acute Stroke.

OBJECTIVE: To test the hypothesis that cognitive demands influence motor performance during recovery from acute stroke, we tested patients with acute stroke on 2 motor tasks with different cognitive demands and related task performance to cognitive impairment and neuroanatomic injury. METHODS: We assessed the contralesional and ipsilesional upper extremities of a cohort of 50 patients with weakness after unilateral acute ischemic stroke at 3 time points with 2 tasks: the Box & Blocks Test, a task with greater cognitive demand, and Grip Strength, a simple and ballistic motor task. We compared performance on the 2 tasks, related motor performance to cognitive dysfunction, and used voxel-based lesion symptom mapping to determine neuroanatomic sites associated with motor performance. RESULTS: Consistent across contralesional and ipsilesional upper extremities and most pronounced immediately after stroke, Box & Blocks scores were significantly more impaired than Grip Strength scores. The presence of cognitive dysfunction significantly explained up to 33% of variance in Box & Blocks performance but was not associated with Grip Strength performance. While Grip Strength performance was associated with injury largely restricted to sensorimotor regions, Box & Blocks performance was associated with broad injury outside sensorimotor structures, particularly the dorsal anterior insula, a region known to be important for complex cognitive function. CONCLUSIONS: Together, these results suggest that cognitive demands influence upper extremity motor performance during recovery from acute stroke. Our findings emphasize the integrated nature of motor and cognitive systems and suggest that it is critical to consider cognitive demands during motor testing and neurorehabilitation after stroke.

Endoscopic Versus Open Total Vault Reconstruction of Sagittal Craniosynostosis.

Sagittal craniosynostosis is the most common form of congenital cranial deformity. Surgical interventions are performed either open or endoscopic. Advancements in minimally invasive surgery have enabled the development of the endoscopic suturectomy technique. This is contrasted to the traditional open cranial vault reconstruction. There is a paucity of data comparing the head shape changes from both techniques. This study aims to compare the morphological outcome of endoscopic suturectomy versus total cranial vault reconstruction. METHODS: This is a retrospective comparative study involving 55 cases of sagittal craniosynostosis, 37 of which has open total cranial vault reconstruction and 18 had endoscopic suturectomy procedure. Preoperative and postoperative 3D photographs of both groups were analyzed and compared. The change in correction between preoperative and postoperative state was measured against a crowd-driven standard for acceptable head shape. RESULTS: Total cranial vault had higher percentage change between pre and postoperative cranial index than endoscopic suturectomy (14.7% versus 7.7%, P = 0.003). However, both techniques were able to achieve the minimum standard of 70% correction (TCV 107.5%, ES 100.4%, P = 0.02). CONCLUSION: Total cranial vault and endoscopic suturectomy are effective in correcting scaphocephaly among children with sagittal craniosynostosis. Additionally, both techniques are able to achieve a percentage correction that exceeds the 70% benchmark established by the lay public.

Hormone receptor expression and outcomes in low-grade serous ovarian carcinoma.

OBJECTIVE: Low-grade serous ovarian carcinomas (LGSC) are frequently ER/PR positive, though the mechanisms by which ER/PR regulate prognosis or anti-estrogen treatment efficacy are poorly understood. We studied ER/PR expression in LGSC tumors and cell lines to evaluate patient outcomes and cellular treatment responses. METHODS: LGSC tumors and patient-derived cell lines were studied from patients with advanced-stage (III/IV) disease. Tumor samples and clinical data were obtained from the Canadian Ovarian Experimental Unified Resource (COEUR-tissue microarray) and the Ovarian Cancer Research (OvCaRe) tissue bank. ER/PR expression was assessed by both Western blot and immunohistochemistry (IHC). Two different IHC scoring systems (simple and Allred) were used. Cox regression was used to identify factors (age, disease residuum, ER/PR status, etc.) associated with progression-free (PFS) and overall survival (OS). Estradiol and tamoxifen proliferation and viability experiments were performed in LGSC cell lines. RESULTS: In 55 LGSC cases studied, median follow-up was 56 months (range 1-227). Fifty-three (96%) cases strongly expressed ER whereas 37 (67%) expressed PR. Cox-regression analysis showed that residuum (p < 0.001) was significantly associated with PFS, whereas both ER Allred score (p = 0.005) and residuum (p = 0.004) were significant for OS. None of the LGSC cell lines expressed PR. Loss of PR and ER expression over time was detected in LGSC tumors and cell lines respectively. Estrogen and tamoxifen treatment did not alter LGSC cell proliferation or viability in-vitro. CONCLUSIONS: In patients with advanced LGSC, higher ER Allred scores were significantly associated with better overall survival. ER/PR expression changed over time in both LGSC tumors and cell lines. Better translational research models are needed to elucidate the molecular mechanisms of ER/PR signalling in LGSC.

Use of plasma DNA to predict mortality and need for intensive care in patients with abdominal pain.

BACKGROUND: We investigated the value of plasma deoxyribonucleic acid concentrations in patients presenting with acute abdominal pain to predict need for intensive care or mortality. METHODS: Plasma deoxyribonucleic acid taken from patients with acute abdominal pain was analyzed for the beta-globin gene using the quantitative polymerase chain reaction. The primary outcome measure was the combined 28-day mortality or admission to the intensive care unit. RESULTS: Of 287 consecutive patients with acute abdominal pain recruited, 12 patients were admitted to the intensive care unit and/or died. Median plasma DNA concentrations were higher in patients with cancer and major organ inflammation. Mean plasma DNA concentrations were three-fold higher in patients with systemic inflammatory response syndrome, five-fold higher in patients who died within 28 days, and eight-fold higher in patients admitted to the intensive care unit. The area under the receiver operator curve for plasma DNA concentrations and intensive care unit admission/mortality was 0.804. At a cut-off of 1100 GE/ml, the sensitivity was 67% (95%CI 35-90) and specificity was 89% (95%CI 84-92). At a cut-off of 175 GE/ml, the sensitivity was 100% (95%CI 73-100) and specificity was 30% (95%CI 25-36). Plasma DNA concentration predicted need for intensive care unit admission or death (adjusted odds ratio 1.4; P<0.0001). CONCLUSIONS: Plasma DNA may have a role in patients with acute abdominal pain as a marker for inflammation and cancer, and a predictor of intensive care unit admission/mortality.

Comparison of plasma beta-globin DNA and S-100 protein concentrations in acute stroke.

BACKGROUND: This study aimed to compare changes in plasma beta-globin DNA and serum S100 protein to diagnose stroke and for predicting mortality and morbidity. METHODS: Patients with stroke-like symptoms presenting to the emergency department of a Hong Kong hospital were recruited. Plasma DNA was analyzed for the beta-globin gene with fluorescent-based PCR. S100 concentrations were determined using ELISA. Primary outcomes were diagnosis of stroke, mortality, and modified Rankin Score (mRS) after 6 months. RESULTS: One hundred ninety-seven consecutive patients recruited, 118 (60%) ischemic stroke, 35 (18%) hemorrhage and 44 (22%) with no acute neuroimaging changes. Serum S100 and plasma DNA were increased in 126 (p<0.0010) and 36 (p=0.21) stroke patients respectively vs. controls. Median plasma DNA was higher in hemorrhagic stroke than those without (1725 vs. 1050 kilogenome-equivalents/l, p=0.0104). Median plasma DNA was higher in mRS>2 vs. mRS2 patients vs. mRS

Circulating nucleic acids and critical illness.

This article reviews some of the early work that has been performed to investigate the potential roles of circulating nucleic acids as prediction markers in acute illness and injury. Circulating DNA and RNA concentrations are elevated early in patients with trauma, stroke and ACS, and are generally highest in patients with a high risk of death. Circulating nucleic acids may be useful markers for the evaluation and risk-stratification of such patients.

Plasma beta-globin DNA as a prognostic marker in chest pain patients.

BACKGROUND: Acute coronary syndrome may involve cell death and the release of nucleic acids into the circulation. We thus investigated whether plasma DNA concentrations are increased and determined its prognostic significance in patients with ACS. METHODS: Real-time polymerase chain reaction was used to quantitatively measure the beta-globin gene from blood samples taken from patients presenting to an emergency department with chest pain of probable cardiac cause. RESULTS: Samples from 58 patients with chest pain, and from 21 age- and sex-matched healthy control subjects were analysed. Compared with the control group, median plasma DNA concentrations were increased 1.5-fold in patients with minor cardiac injury, were increased further in patients with STEA and STEMI, and were the highest in those patients who died within 2 years (P=0.0005; post-hoc Dunn's, P<0.05). Median plasma DNA concentrations were higher in patients who later developed heart failure (1060 vs. 500 kGE/l; P=0.0095); higher in patients who later reinfarcted (1000 vs. 530 kGE/l; P=0.0298); higher in patients who had a cardiac arrest in that admission (1350 vs. 525 kGE/l; P=0.04); and were higher in patients who were readmitted within 6 months of discharge (725 vs. 475 kGE/l; P=0.04). CONCLUSION: Plasma DNA is a potential marker for post-ACS complications.

Plasma DNA as a prognostic marker for stroke patients with negative neuroimaging within the first 24 h of symptom onset.

Modern neuroimaging safely and reliably diagnoses stroke and provides information for outcome prediction. However, some patients with clinical stroke have no detectable abnormality on neuroimaging and other patients are not fit for such investigations. Therefore, we evaluated the potential of plasma DNA and serum S100 protein concentrations to predict post-stroke mortality and morbidity in patients with negative neuroimaging results. Patients with stroke-like symptoms but negative neuroimaging results were recruited. Both plasma and serum were collected from each patient for plasma DNA and serum S100 analysis. The primary outcome measures were 6-month mortality and morbidity using the post-stroke modified Rankin score (mRS). Forty-four patients were recruited to the study. Seventeen (39%) patients were classified as post-stroke mRS grades 3-6. The median plasma DNA concentration of this group of patients was significantly higher than that of patients with post-stroke mRS grades 0-2. Median serum S100 protein concentrations did not show significant differences between the two groups. Plasma DNA concentrations > 800 kilogenome-equivalent/l have a sensitivity of 42% and a specificity of 100% for predicting 6-month post-stroke mRS (grades 0-2), with an area under the receiver operator characteristic (ROC) curve of 0.742. By comparison, serum S100 protein concentrations > 0.09 microg/l have a sensitivity of 48% and specificity of 75% for predicting 6-month post-stroke mRS (grades 0-2), and the area under the curve is 0.542. Plasma DNA concentration predicts post-stroke morbidity and mortality in patients with negative neuroimaging, and may be more effective than S100 protein measurement.

Time course of early and late changes in plasma DNA in trauma patients.

BACKGROUND: Cell-free DNA concentrations increase in the circulation of patients after trauma and may have prognostic potential, but little is know concerning the temporal changes or clearance of the DNA or its relationships with posttraumatic complications. We investigated temporal changes in plasma DNA concentrations in patients after trauma with use of real-time quantitative PCR. METHODS: Serial plasma samples were taken from two trauma populations. In the first study, samples were collected every 20 min from 25 patients within the first 3 h of trauma. In the second study, samples were collected every day from 36 other trauma patients admitted to the intensive care unit (ICU). RESULTS: In the first study, plasma DNA was increased within 20 min of injury and was significantly higher in patients with severe injury and in patients who went on to develop organ failure. In patients with less severe injuries, plasma DNA concentrations decreased toward reference values within 3 h. In the second study, plasma DNA concentrations were higher in patients who developed multiple organ dysfunction syndrome between the second and fourth days of admission than in patients who did not develop the syndrome. In patients who remained in the ICU with continuing organ dysfunction, plasma DNA remained higher than in healthy controls even at 28 days after injury. Most survivors with multiple organ dysfunction syndrome showed an initial very high peak followed by a prolonged smaller increase. CONCLUSIONS: Plasma DNA concentrations increase early after injury and are higher in patients with severe injuries and in those who develop organ failure. Increased plasma DNA persists for days after injuries, especially in patients with multiple organ dysfunction syndrome.

Quantitative analysis of circulating mitochondrial DNA in plasma.

BACKGROUND: Recent studies have demonstrated the existence of circulating mitochondrial DNA in plasma and serum, but the concentrations and physical characteristics of circulating mitochondrial DNA are unknown. The aim of this study was to develop an assay to quantify mitochondrial DNA in the plasma of healthy individuals. METHODS: We adopted a real-time quantitative PCR approach and evaluated the specificity of the assay for detecting mitochondrial DNA with a cell line (rho(0)) devoid of mitochondria. The concentrations and physical characteristics of circulating mitochondrial DNA were investigated by experiments conducted in three modules. In module 1, we evaluated the concentrations of mitochondrial DNA in plasma aliquots derived from four blood-processing protocols. In module 2, we investigated the existence of both particle-associated and free forms of mitochondrial DNA in plasma by subjecting plasma to filtration and ultracentrifugation. In module 3, we used filters with different pore sizes to investigate the size characteristics of the particle-associated fraction of circulating mitochondrial DNA. RESULTS: The mitochondrial DNA-specific, real-time quantitative PCR had a dynamic range of five orders of magnitude and a sensitivity that enabled detection of one copy of mitochondrial DNA in plasma. In module 1, we found significant differences in the amounts of circulating mitochondrial DNA among plasma aliquots processed by different methods. Data from module 2 revealed that a significant fraction of mitochondrial DNA in plasma was filterable or pelletable by ultracentrifugation. Module 3 demonstrated that filters with different pore sizes removed mitochondrial DNA from plasma to different degrees. CONCLUSIONS: Both particle-associated and free mitochondrial DNA are present in plasma, and their respective concentrations are affected by the process used to harvest plasma from whole blood. These results may have implications in the design of future studies on circulating mitochondrial DNA measured in different disease conditions.

Prognostic use of circulating plasma nucleic acid concentrations in patients with acute stroke.

BACKGROUND: At present there is no simple, accurate blood test that may be used to determine the severity of stroke or to predict mortality and morbidity in stroke patients presenting to emergency departments. METHODS: Patients with stroke-like symptoms who presented to an emergency department of a university hospital in Hong Kong were recruited for the study. DNA extracted from patients' plasma was analyzed for the beta-globin gene with a fluorescent-based PCR test. The primary outcome measures were in-hospital and 6-month mortality and morbidity using the post-stroke modified Rankin Score. RESULTS: Among the 88 consecutive patients recruited to the study, 70 (80%) had ischemic stroke, 11 (13%) had intracerebral hemorrhage, and 7 (8%) had transient ischemic attacks. Median plasma DNA concentrations taken within 3 h of symptom onset were higher in patients who died compared with those who survived at discharge (6205 vs 1334 kilogenome-equivalents/L; P = 0.03). Among patients with NIH Stroke Scale scores >8, median plasma DNA concentrations were higher in patients who died compared with those who survived to 6 months (2273 vs 968 kilogenome-equivalents/L; P = 0.002). Plasma DNA concentrations correlated with the volume of cerebral hematoma (r = 0.66; P = 0.03). Plasma DNA concentrations >1400 kilogenome-equivalents/L had a sensitivity of 100% and a specificity of 74.4% for predicting hospital mortality after stroke, and the area under the ROC curve was 0.89 (95% confidence interval, 0.80-0.94). The adjusted odds ratio for plasma DNA concentrations predicting 6-month mortality was 1.6 (1.1-2.4; P = 0.03) and for predicting 6-month post-Rankin Score >2 was 1.8 (1.0-3.3; P = 0.05). CONCLUSION: Plasma DNA concentrations correlate with stroke severity and may be used to predict mortality and morbidity in the emergency room.

Effect of stress hormones on the expression of fibrinogen-binding receptors in platelets.

Acute coagulopathy is a common clinical complication after trauma, and contributes to posttraumatic multiple organ failure. The phenomenon may be due to the effect of stress hormones on platelet adhesion molecule expression after trauma. Catecholamine levels correlate with injury severity scores and changes of L-selectin expression on leucocytes, whilst adrenaline (ADR) (epinephrine) alone also activates platelets. This study thus investigates the effects of ADR and noradrenaline (NOR) (norepinephrine) on platelets, at doses similar to those found in the plasma of normal and trauma subjects. Blood was taken from 19 healthy subjects and placed in tubes containing sodium citrate. Anti-platelet-bound fibrinogen monoclonal antibody was used to identify the activated platelets while anti-CD41 was used to identify platelets with and without activation. Five increasing concentrations of ADR and NOR (1, 3, 5, 10, 30 nmol/l) as well as one negative control (0.9% normal saline) and one positive control (10 micromol/l adenosine diphosphate/ADP) were prepared for the stimulation. A whole blood protocol was used in order to minimize any activation artefacts, which might be created by centrifugation. The percentage of platelets expressing fibrinogen receptors increased significantly with ADR and NOR even at the lowest dose (1 nmol/l) and continued to increase in a dose-dependent manner. Although the effect of ADR was greater than NOR in stimulating platelets to express fibrinogen receptors, the average number of fibrinogen receptors on each platelet was constant. ADR and NOR activated platelets to express fibrinogen receptors at doses that are similar to those found in the plasma of trauma patients.

Presence of filterable and nonfilterable mRNA in the plasma of cancer patients and healthy individuals.

BACKGROUND: As RNA is labile, we investigated whether circulating RNA in human plasma may be present in a particle-associated form. METHODS: Blood was collected from 27 healthy individuals and 16 hepatocellular carcinoma (HCC) patients. The plasma from each individual was processed by two means: filtration through filters with different pore sizes (from 5 microm to 0.22 microm) and ultracentrifugation. We assessed plasma RNA content by a real-time quantitative reverse transcription-PCR assay for glyceraldehyde-3-phosphate dehydrogenase (GAPDH) transcripts and plasma DNA by a real-time quantitative PCR assay for the beta-globin gene. RESULTS: The plasma GAPDH mRNA concentrations in the healthy individuals were significantly different in every pair of these filter sizes (P <0.05 for each pair). Overall, the plasma GAPDH mRNA concentration was higher by a median of 15-fold (interquartile range, 10- to 24-fold) in the paired unfiltered sample than in the sample filtered through a 0.22 microm filter. In contrast, no significant difference was seen in beta-globin DNA concentrations among different pore-size-filtered plasma samples (P = 0.455). Similarly, a significant difference was observed for RNA, but not DNA, between unfiltered plasma and ultracentrifuged plasma (P <0.05). No significant difference in GAPDH mRNA concentrations was seen between the 0.22-microm-filtered plasma samples and the ultracentrifuged plasma samples (P >0.05). In HCC patients, filtration with a 0.22 microm filter produced a median 9.3-fold (interquartile range, 6.9- to 311-fold) reduction in GAPDH mRNA concentration in plasma. Plasma GAPDH mRNA concentrations in HCC patients were significantly higher than those in healthy individuals, both with or without filtration (P <0.0 5 for filtered plasma samples; P <0.005 for unfiltered plasma samples). CONCLUSIONS: A substantial proportion of plasma mRNA species is particle-associated. In HCC patients, both circulating particle- and non-particle-associated plasma RNA are increased.