Pervasive nuclear envelope ruptures precede ECM signaling and disease onset without activating cGAS-STING in Lamin-cardiomyopathy mice.

Nuclear envelope (NE) ruptures are emerging observations in Lamin-related dilated cardiomyopathy, an adult-onset disease caused by loss-of-function mutations in Lamin A/C, a nuclear lamina component. Here, we test a prevailing hypothesis that NE ruptures trigger the pathological cGAS-STING cytosolic DNA-sensing pathway using a mouse model of Lamin cardiomyopathy. The reduction of Lamin A/C in cardio-myocyte of adult mice causes pervasive NE ruptures in cardiomyocytes, preceding inflammatory transcription, fibrosis, and fatal dilated cardiomyopathy. NE ruptures are followed by DNA damage accumulation without causing immediate cardiomyocyte death. However, cGAS-STING-dependent inflammatory signaling remains inactive. Deleting cGas or Sting does not rescue cardiomyopathy in the mouse model. The lack of cGAS-STING activation is likely due to the near absence of cGAS expression in adult cardiomyocytes at baseline. Instead, extracellular matrix (ECM) signaling is activated and predicted to initiate pro-inflammatory communication from Lamin-reduced cardiomyocytes to fibroblasts. Our work nominates ECM signaling, not cGAS-STING, as a potential inflammatory contributor in Lamin cardiomyopathy.

The cGAS-STING pathway is dispensable in a mouse model of LMNA-cardiomyopathy despite nuclear envelope rupture.

Mutations in the nuclear Lamin A/C gene (LMNA) cause diverse degenerative disorders, including malignant dilated cardiomyopathy in adults. A prevailing hypothesis postulates that LMNA mutations cause nuclear envelope ruptures that trigger pathogenic inflammatory signaling via the cGAS-STING cytosolic DNA-sensing pathway. Here, we provide evidence against this hypothesis, using a mouse model of LMNA-related cardiomyopathy that mimics Lamin A/C protein reduction observed in patient cardiomyocytes. We observed that pervasive nuclear envelope ruptures preceded the onset of cardiac transcriptional modulation and dilated cardiomyopathy. Nuclear ruptures activated DNA damage response without causing immediate cardiomyocyte death. However, cGAS-STING downstream cytokine genes remained inactive in the mutant cardiomyocytes. Deleting cGas or Sting did not alleviate cardiomyopathy. Instead, extracellular matrix signaling was predicted to emanate from Lamin A/C-reduced cardiomyocytes to communicate with fibroblasts in the heart. These findings suggest that cGAS-STING is not a major pathogenetic contributor to LMNA-related dilated cardiomyopathy in adult humans.

Risk of Colorectal Cancer in Serrated Polyposis Syndrome: A Systematic Review and Meta-analysis.

BACKGROUND & AIMS: Serrated polyposis syndrome (SPS) is characterized by development of numerous serrated lesions throughout the colorectum and increased risk of colorectal cancer (CRC). However, SPS has been an underrecognized CRC predisposition syndrome, and the true risk of CRC in SPS, both overall and in surveillance, is not known. The aim of this systematic review and meta-analysis is to describe the risk of CRC in patients with SPS. METHODS: Electronic databases were searched on March 25, 2021, for studies describing CRC risk in SPS. Random-effects meta-analysis was performed to assess pooled risk of CRC among SPS patients. Primary outcomes were risk of CRC at time of SPS diagnosis and during surveillance following diagnosis of SPS. Secondary outcomes included risk of CRC prior to diagnosis of SPS and effect of World Health Organization subtype on CRC risk. RESULTS: Thirty-six studies including 2788 patients with SPS were included in the analysis. Overall risk of CRC in SPS was 19.9% (95% confidence interval [CI], 15.3%-24.5%). CRC risk at the time of diagnosis was 14.7% (95% CI, 11.4%-18.8%), while risk during surveillance was 2.8% (95% CI, 1.8%-4.4%), or 7 cases per 1000 person-years. SPS patients also had a high incidence of history of CRC prior to SPS diagnosis (7.0%; 95% CI, 4.6%-11.7). Subgroup analysis did not reveal any significant differences based on World Health Organization subtype. CONCLUSIONS: Our meta-analysis demonstrated that patients with SPS have an elevated risk of CRC, which is highest at the time of diagnosis and suggests the importance of early SPS recognition and screening to modify CRC risk. The persistently elevated CRC risk during surveillance supports current guidelines recommending heightened surveillance protocols.

Target specific serologic analysis of COVID-19 convalescent plasma.

This study compared the performance of four serology assays for Coronavirus Disease 2019 (COVID-19) and investigated whether COVID-19 disease history correlates with assay performance. Samples were tested at Northshore using the Elecsys Anti-SARS-CoV-2 (Roche Diagnostics), Access SARS-CoV-2 IgG anti-RBD (Beckman Coulter), and LIAISON SARS-CoV-2 S1/S2 IgG (DiaSorin) as well as at Genalyte using Maverick Multi-Antigen Serology Panel. The study included one hundred clinical samples collected before December 2019 and ninety-seven samples collected from convalescent plasma donors originally diagnosed with COVID-19 by PCR. COVID-19 disease history was self-reported by the plasma donors. There was no difference in specificity between the assays tested. Clinical sensitivity of these four tests was 98% (Genalyte), 96% (Roche), 92% (DiaSorin), and 87% (Beckman). The only statistically significant differences in clinical sensitivity was between the Beckman assay and both Genalyte and Roche assays. Convalescent plasma donor characteristics and disease symptoms did not correlate with false negative results from the Beckman and DiaSorin assays. All four tests showed high specificity (100%) and varying sensitivities (89-98%). No correlations between disease history and serology results were observed. The Genalyte Multiplex assay showed as good or better sensitivity to three other previously validated assays with FDA Emergency Use Authorizations.

Rapidly Establishing a Hospital-Based Convalescent Plasma Collection Center With the Alyx Apheresis Collection Device.

The effort to collect convalescent plasma from individuals who recovered from COVID-19 began in earnest during the spring of 2020. Either whole blood or apheresis donations were obtained, the latter yielding higher numbers of units per donor per collection and more frequent collections. The NorthShore University HealthSystem blood donor center purchased 2 Alyx (Fresenius Kabi) apheresis plasma collection devices and quickly implemented them in order to collect COVID-19 convalescent plasma. Apheresis-experienced and inexperienced phlebotomists operated the instruments. Donors were collected >14 days from symptom resolution and all donors were negative by SARS-CoV-2 nasopharyngeal swab. Both internal metrics of performance as well as a post donation survey were used to evaluate the feasibility implementing this collection program. During the first 100 days of the collection program, 650 plasma units were collected. In particular, during the first week of the program, 38 units were collected and distributed to hospitals under the emergency investigational new drug and expanded access program. Fifty-one donors (15%) were deferred due to vital signs out of range or donor screening questions. Thirty-one donors (10%) were deferred due to positive nasopharyngeal swab. Lower than target yield occurred in 16.6% of collections due to donor reactions or flow errors. Donors rated the overall program lower, but not the staff, when they reported symptoms related to collection. In conclusion, a hospital-based apheresis convalescent plasma collection program can be rapidly implemented. Donor reaction rates and vein infiltration rates should be carefully monitored for each phlebotomist.

Persistence of SARS-CoV-2 nasopharyngeal swab PCR positivity in COVID-19 convalescent plasma donors.

BACKGROUND: Nucleic acid persists after symptom resolution and infectivity for many viral infections via delayed clearance of nucleic acid fragments, non-infectious particles, or transmissible virus. For Coronavirus Disease 2019 (COVID-19), the relationship between nasopharyngeal (NP) swab positivity, the development of antibodies against COVID-19, and clinical history are unclear. STUDY DESIGN AND METHODS: Individuals who recovered from COVID-19 and volunteered to donate convalescent plasma (CP) were screened by NP swab PCR, responded to a questionnaire, and were tested for anti-COVID-19 antibodies. RESULTS: A proportion of 11.8% of individuals tested positive for SARS-CoV-2 by NP swab PCR greater than 14 days after the resolution of symptoms of active disease, including one donor who had asymptomatic disease and tested positive by NP swab 41 days after her initial diagnosis. Clinical history did not show a significant correlation with persistence of NP swab positivity. Also, NP swab positivity >14 days from symptom resolution did not correlate with anti-COVID-19 serology results. IgG anti-SARS-CoV-2 spike antibody strength correlated with hospitalization for COVID-19 using two different assays. Total anti-SARS-CoV-2 nucleocapsid antibody strength correlated with time from symptom resolution to sample collection and symptom duration. CONCLUSIONS: SARS-CoV-2 nucleic acid is detectable long after the resolution of symptoms in a significant percentage of previously diagnosed individuals, which is important to consider when interpreting PCR swab results. Persistence of PCR positivity does not correlate with antibody strength or symptoms of COVID-19. If anti-spike antibody is used to assess CP potency, individuals who suffered severe COVID-19 disease symptoms may represent better donors.

Is Next-Generation Sequencing Alone Sufficient to Reliably Diagnose Gliomas?

The power and widespread use of next-generation sequencing (NGS) in surgical neuropathology has raised questions as to whether NGS might someday fully supplant histologic-based examination. We therefore sought to determine the feasibility of relying on NGS alone for diagnosing infiltrating gliomas. A total of 171 brain lesions in adults, all of which had been analyzed by GlioSeq NGS, comprised the study cohort. Each case was separately diagnosed by 6 reviewers, based solely on age, sex, tumor location, and NGS results. Results were compared with the final integrated diagnoses and scored on the following scale: 0 = either wrong tumor type or correct tumor type but off by 2+ grades; 1 = off by 1 grade; 2 = exactly correct. Histology alone was treated as a seventh reviewer. Overall reviewer accuracy ranged from 81.6% to 94.2%, while histology alone scored 87.1%. For glioblastomas, NGS was more accurate than histology alone (93.8%-97.9% vs 87.5%). The NGS accuracy for grade II and III astrocytoma and oligodendroglioma was only 54.3%-84.8% and 34.4%-87.5%, respectively. Most uncommon gliomas, including BRAF-driven tumors, could not be accurately classified just by NGS. These data indicate that, even in this era of advanced molecular diagnostics, histologic evaluation is still an essential part of optimal patient care.

Equol, a Metabolite of Daidzein, Is More Efficient than Daidzein for Bone Formation in Growing Female Rats.

Few studies have examined the effects of isoflavones and particularly equol, a metabolite of the isoflavone daidzein, on bone formation during the growth period in animals. The present study investigated the effects of orally administered daidzein or equol on bone formation and bone mineral density in growing female rats. Female Sprague-Dawley rats, aged 3 weeks, were divided into four groups (n = 8 per group) as follows: rats were orally administered a corn oil, 8 mg/day of daidzein, 4 mg/day of equol or 8 mg/day of equol in corn oil for 4 weeks. Daidzein and equol increased the bone mineral density of growing female rats by stimulating bone formation without exhibiting a substantial effect on the weight of their reproductive organs. Bone growth caused by increased bone mineralizing surface and bone formation rate in rats administered with equol was approximately twice that of rats administered with daidzein. These results suggest that equol might be more efficient than daidzein for bone formation in growing female rats. Copyright © 2015 John Wiley & Sons, Ltd.

Effect of high beta-glucan barley on serum cholesterol concentrations and visceral fat area in Japanese men--a randomized, double-blinded, placebo-controlled trial.

This study investigated whether the consumption of a diet in which high-beta-glucan barley replaced rice would reduce the visceral fat area as well as the serum low-density lipoprotein cholesterol (LDL-C) and total cholesterol (TC) in hypercholesterolemic Japanese men. A randomized, double-blinded, placebo-controlled intervention study was conducted in 44 hypercholesterolemic Japanese men with a body mass index (BMI) >22 kg/m2. The subjects were randomly assigned to groups consuming either rice (placebo group) or a mixture of rice and pearl barley with a high beta-glucan content (test group, 7.0 g beta-glucan per day) for 12 weeks. Blood samples were taken, and CT scan obtained before the trial and every four weeks during the trial. The pearl barley intake significantly reduced serum concentrations of LDL-C (P = 0.041) and TC (P = 0.037) during the trial. Significant differences between the test and placebo groups were found for the visceral fat (P = 0.039), BMI (P = 0.015), and waist circumference (P = 0.011) at the end point. The consumption of pearl barley with a high beta-glucan content reduces not only LDL-C but also visceral fat area.

Differential effects of isoflavones on bone formation in growing male and female mice.

Few studies have examined the effects of isoflavones on bone formation during growth period in male and female animals. In this study, the effects of daidzein or genistein on bone formation were assessed in immature male and female mice. Five-week-old male and female mice were divided respectively into 3 groups (n=8 per group) as follows: control group (C) fed a control diet (AIN-93G), daidzein group (D) fed a control diet containing 0.08% pure daidzein, and genistein group (G) fed a control diet containing 0.08% pure genistein. After 4 weeks, the male D group had a significantly higher bone mineral density (BMD) in whole body, lumbar spine, and femur than did the C group. On the contrary, BMD of the whole body and femur in the female D group was significantly lower than that in the C group. The BMD of the whole femur in the male G group also was significantly higher than for the C group. Histologic analysis revealed that the bone formation rate was significantly higher in the male D and G groups, and lower in the female D group compared with the C group. These results suggest that daidzein has a specific, sexually dimorphic effect on bone formation and BMD during growth period in mice.

A simplified modification of the AOAC official method for determination of total dietary fiber using newly developed enzymes: preliminary interlaboratory study.

A preliminary interlaboratory study was conducted to evaluate the validity of the modified AOAC method for determination of total dietary fiber by Tada and Innami, in which the 3-step enzymatic digestion process in AOAC Method 991.43 is modified to a 2-step process without pH adjustment. Total dietary fiber contents in 8 representative foodstuffs were measured using both the original AOAC Method 991.43 and the modified method in 6 research facilities in Japan. Repeatability relative standard deviations, reproducibility relative standard deviations, and Horwitz ratio values from the modified method were equivalent to those from AOAC Method 991.43, except in the rice sample. However, this exceptional case shown in the modified method was entirely dissolved by the addition of alpha-amylase stabilizing agents. The modified method, which shortens the process of enzymatic digestion from 3 to 2 steps and in which only reaction temperature is adjusted under the same pH, was found not only to give accurate values comparable to the original method, but also to substantially reduce the labor required by the laboratory staff in the process of routine analysis. This study revealed that the validity of the modified method was further ensured by adding alpha-amylase stabilizing agents to the reaction system.

Effects of genistein, an isoflavone, on pregnancy outcome and organ weights of pregnant and lactating rats and development of their suckling pups.

There is a general agreement that isoflavones can be beneficial to health in adults. However, isoflavones are well known as endocrine-disrupting chemicals. It should be considered that soy foods might adversely affect the reproductive system and infants. The aim of this study was to evaluate the effects of genistein, an isoflavone, on dams and their offspring. Maternal rats were fed diets containing genistein at levels of 0 and 0.5 g/kg diet from pregnancy day 5 to postnatal day 13. No effects of genistein on the delivery, anogenital distance, reproductive organ weight, and body weight of the infants at birth were observed. There were no consistent effects on suckling pups after continuous genistein exposure during their fetal and suckling stages through their mothers, and there was no difference in effects according to the periods of exposure during pregnancy and lactation. We also observed no significant effect on the growth of offspring after weaning. Moreover, while we observed that the serum concentration of triiodothyronine (T3) in dams decreased, the result was a tendency, not a significant decrease. Our study suggested that maternal ingestion of genistein might have not induced serious adverse effects on dams, fetuses, infants or offspring during growth. However, the results indicated in many papers suggest the necessity of further study on the safety of genistein.

High-fat diet enhances the accumulation of hexachlorobenzene (HCB) by pregnant rats during continuous exposure to HCB.

To investigate the influence of a high-fat diet on HCB distribution and accumulation, pregnant rats in study 1 were fed a high-fat or control diet containing HCB, and, in study 2, pregnant rats were given a single HCB dose by intragastric gavage and HCB-free high-fat or control diet. In study 1, the high-fat diet group had higher HCB concentrations in fat tissues and liver than did the controls. In study 2, although the total amounts of HCB in the fat tissue and liver were greater in the high-fat diet group than in the controls, no significant differences in HCB concentration were observed between the two groups. The high-fat diet group also showed more fecal excretion of HCB. Therefore, HCB accumulation in rats fed a high-fat diet was enhanced more by continuous exposure to HCB than by administration of a single dose.

Increased incidence rate of colorectal tumors due to the intake of a soluble dietary fiber in rat chemical carcinogenesis can be suppressed by substituting partially an insoluble dietary fiber for the soluble one.

In epidemiologic studies on human colorectal tumors, results on the relative protective effect of soluble and insoluble fibers are not consistent. We studied in this work the effect in rats of feeding guar gum or guar gum together with cellulose on the incidence of colorectal tumors induced by 1,2-dimethylhydrazine. The results were as follows: (i) The enhancement of tumor formation by feeding solely guar gum (guar gum group) was suppressed completely when two-thirds of the guar gum was replaced with cellulose (cellulose-guar gum group). The odds ratio for tumor formation was 0.075 (95% CI 0.006-0.936, p = 0.044) for guar gum group vs. no fiber control and 0.833 (0.134-5.167, p = 0.83) for cellulose-guar gum group vs. the control. (ii) In both groups, serum cholesterol and triglyceride levels decreased significantly compared to the no fiber control group, and fecal excretion of total bile acids almost doubled. (iii) In guar gum group rats, the deconjugation activity (beta-glucuronidase, beta-glucosidase) was higher than the control or cellulose-guar gum group rats. (iv) The amount of cecal short-chain fatty acids was almost double in guar gum group rats compared to the cellulose-guar gum group or the control rats, and pH of the cecal content of the guar gum group rats had a tendency to be lower. (v) The concentration of fecal secondary bile acids was extremely low in the younger rats of the guar gum group. From these results, it seemed significant to study the cancer preventive effect of the mixed feeding to experimental animals of water-soluble and insoluble fibers instead of the singular feeding.

Effects of geranylgeranoic acid in bone: induction of osteoblast differentiation and inhibition of osteoclast formation.

Retinoids are known to be of special importance for normal bone growth and development. Recently, we reported that retinoids not only induced osteoblast differentiation, but also inhibited osteoclast formation in vitro. In this study, we examined the osteogenic effects of geranylgeranoic acid (GGA), a chemically synthesized acyclic retinoid, in bone in vitro and in vivo. GGA not only suppressed proliferation of osteoblastic MC3T3-E1 cells, but also up-regulated differentiation markers of osteoblasts such as alkaline phosphatase (ALP) activity and expression of osteopontin (OP) messenger RNA (mRNA). In contrast, GGA inhibited osteoclast formation induced by 1alpha,25-dihydroxyvitamin D3 [1alpha,25(OH)2D3] in cocultures of mouse bone marrow cells and primary osteoblasts. Treatment of stromal ST2 cells with GGA restored the 1alpha,25(OH)2D3- or prostaglandin E2 (PGE2)-induced suppression of osteoprotegerin (OPG) mRNA expression. GGA inhibited osteoclast formation induced by macrophage colony-stimulating factor (M-CSF) and soluble receptor activator of nuclear factor kappaB ligand (sRANKL) in the culture of bone marrow macrophages. Thus, it is likely that GGA inhibits osteoclast formation by affecting both osteoblasts and osteoclast progenitors in the coculture system. Furthermore, in vivo, GGA increased bone mineral density (BMD) of total as well as distal femur in a P6 strain of senescence-accelerated mice (SAMP6). These results indicate that GGA increases bone mass by maintaining a positive balance of bone turnover by inducing osteoblast differentiation and suppressing osteoclast formation.