Abstract 290: Lower Limb Revascularization of Patients with Peripheral Artery Disease Reduces Human Enterovirus Infection of the Gastrocnemius

2015 ◽  
Vol 35 (suppl_1) ◽  
Author(s):  
Julian Kim ◽  
George Casale ◽  
Zhen Zhu ◽  
Stanley Swanson ◽  
Panagiotis Koutakis ◽  
...  
Keyword(s):  
Copy Number ◽  
Viral Rna ◽  
Human Enterovirus ◽  
Peripheral Artery ◽  
Copy Numbers ◽  
Viral Copy Number ◽  
Viral Copy ◽  
Wet Weight ◽  
Artery Disease ◽  
Ischemic Muscle

Introduction: Peripheral artery disease (PAD), caused by obstruction of arteries supplying the lower extremities, is characterized by leg muscle degeneration. Recently, we showed that infectious human enterovirus (HEV), which has been implicated in other myodegenerative diseases, is present in ischemic muscle of PAD patients and that viral copy numbers correlate with disease severity. Hypothesis: We tested the hypothesis that prevalence of infectious HEV and viral copy number in the gastrocnemius of patients with PAD decrease in response to revascularization. Methods: Gastrocnemius biopsies were collected from controls (N=14) and from PAD patients (N=17), before and after revascularization. Biopsies were examined for the presence of HEV RNA, viral capsid protein, viral RNA copy number, and viral infectivity and for viral sub-genotype. Results: HEV RNA was detected in biopsies of 65% (11/17) of PAD patients and in none of the controls. After revascularization, the gastrocnemius of 5 patients who were HEV positive before surgery no longer harbored detectable virus. The prevalence of HEV infection was reduced from 65% (11/17 patients) before surgery to 35% (6/17 patients) after surgery. Positive-strand viral RNA copy numbers were decreased significantly after revascularization. Mean positive-strand copy number (average 10 2.61 copies/mg muscle wet weight) in post-surgical biopsies that were HEV-positive were lower (p<0.001) compared to mean copy number (average 10 5.46 copies/mg muscle wet weight) in pre-surgical biopsies that were HEV-positive. Copy numbers of negative-strand (template) viral RNA were also decreased after revascularization. Viral replication was confirmed by detection of negative-strand viral RNA in all specimens positive for HEV. Cultures of HeLa and human skeletal muscle cells treated with muscle homogenates showed HEV replication and the presence of HEV capsid protein. By sequence analysis, HEV in PAD muscle exhibited 97% homology with Coxsackievirus B1. Conclusion: Our data introduce HEV infection as a potential mechanism for degeneration of ischemic muscle in PAD patients and demonstrate that revascularization of PAD patients reduces both prevalence of HEV infection and viral copy number in the gastrocnemius.

scholarly journals SARS-CoV-2 Delta variant saliva viral load is 15-fold higher than wild-type strains

2021 ◽  
Author(s):  
Kenichi Imai ◽  
Ryo Ikeno ◽  
Hajime Tanaka ◽  
Norio Takada
Keyword(s):  
Viral Load ◽  
Copy Number ◽  
Null Distribution ◽  
Host Cells ◽  
Wild Type ◽  
Copy Numbers ◽  
Whole Saliva ◽  
Viral Copy Number ◽  
Viral Copy ◽  
Type Strains

The emergence of SARS-CoV-2 Delta variants has escalated COVID-19 cases globally due to their high transmissibility. Since saliva is crucial for SARS-CoV-2 transmission, we hypothesized that a higher viral load of Delta variants in saliva than their parental wild-type strains contributed to the high transmissibility in the first place. However, studies have not reported this particular comparison done with viral copy numbers. Twenty-two genetically confirmed -positive saliva samples for wild-type strain and 32 Delta variants were statistically compared for viral copy number per milliliter determined by real-time qPCR combined with synthesized viral RNA and Poisson's null distribution equation between the groups of wild and variant strains and between whole saliva and centrifugal supernatant in each group. We found that the copy number of the Delta variants was 15.1 times higher than wild-type strains of the whole saliva. In addition, the viral load of both strains in the whole saliva was higher than the pertinent supernatant, indicating that most viruses in the whole saliva are associated with host cells. Meanwhile, more than a million virions per milliliter of the viral load of the variants in the supernatants were 4.0 times higher but not significant than wild-type strains. Humanity must share our findings; the simple but concrete note that Delta variant viral load is abundant in the saliva is critical for preventing the spread of infection.

Infection with antiretroviral-susceptible HIV in an individual adherent to pre-exposure prophylaxis: strategies for treatment initiation

2021 ◽  
pp. 095646242097112
Author(s):  
Jessica M Hughes ◽  
Darrell HS Tan ◽  
Peter Anderson ◽  
Janani Bodhinayake ◽  
Paul A MacPherson
Keyword(s):  
Viral Load ◽  
Copy Number ◽  
Case Reports ◽  
Pharmacy Records ◽  
Treatment Regimens ◽  
Viral Copy Number ◽  
Low Copy Number ◽  
Viral Copy ◽  
Exposure Prophylaxis ◽  
Tenofovir Diphosphate

HIV pre-exposure prophylaxis (PrEP) is effective at preventing sexual acquisition of HIV, and failures in clinical trials are largely attributable to medication nonadherence. We report here a case of infection with a fully susceptible strain of HIV in an individual adherent to PrEP as demonstrated by pharmacy records and intracellular tenofovir diphosphate levels. At diagnosis, the viral load was 90 copies/mL precluding initial genotype testing due to low copy number. While PrEP failure is rare, this case underscores the importance of regular HIV testing for patient on PrEP and prompts discussion regarding the approach to treatment following failure where an initial genotype is not yet available or not possible due to low viral load. Few other case reports of PrEP failure exist in the literature and approaches to treatment varied widely. We suggest the initial viral copy number may guide next steps and discuss the risks and benefits of stopping PrEP, escalating therapy with integrase inhibitors or boosted protease inhibitors, or switching to non-nucleoside antiretroviral treatment regimens.

Activation-Induced CD4+ T Cell Death in HIV-Positive Individuals Correlates with Fas Susceptibility, CD4+ T Cell Count, and HIV Plasma Viral Copy Number

1999 ◽  
Vol 15 (17) ◽  
pp. 1509-1518 ◽  
Author(s):  
D. H. Dockrell ◽  
A. D. Badley ◽  
A. Algeciras-Schimnich ◽  
M. Simpson ◽  
R. Schut ◽  
...  
Keyword(s):  
Cell Death ◽  
T Cell ◽  
Cell Count ◽  
Copy Number ◽  
Cd4 T Cell ◽  
Hiv Positive ◽  
Viral Copy Number ◽  
Viral Copy ◽  
Cd4 T Cell Count

scholarly journals Quantification of SARS-CoV-2 viral copy number in saliva mouthwash samples using digital droplet PCR

2020 ◽  
Author(s):  
Lisa Oberding ◽  
Jia Hu ◽  
Byron Berenger ◽  
Abu Naser Mohon ◽  
Dylan R. Pillai
Keyword(s):  
Viral Load ◽  
Copy Number ◽  
Clinical Samples ◽  
Digital Droplet Pcr ◽  
Viral Copy Number ◽  
Droplet Pcr ◽  
Precise Quantification ◽  
Viral Copy ◽  
Mouth Wash

AbstractSaliva samples were collected through a simple mouth wash procedure and viral load quantified using a technology called digital droplet PCR. Data suggest ddPCR allows for precise quantification of viral load in clinical samples infected with SARS-CoV-2.

Colorimetric loop-mediated isothermal amplification (LAMP) for cost-effective and quantitative detection of SARS-CoV-2: the change in color in LAMP-based assays quantitatively correlates with viral copy number

2021 ◽  
Author(s):  
Everardo González-González ◽  
Itzel Montserrat Lara-Mayorga ◽  
Iram Pablo Rodríguez-Sánchez ◽  
Yu Shrike Zhang ◽  
Sergio O. Martínez-Chapa ◽  
...  
Keyword(s):  
Copy Number ◽  
Quantitative Method ◽  
Diagnostic Performance ◽  
Cost Effective ◽  
Isothermal Amplification ◽  
Quantitative Detection ◽  
Loop Mediated Isothermal Amplification ◽  
Viral Copy Number ◽  
Viral Copy

Colorimetric LAMP for COVID-19 intensified diagnostics: a simple and quantitative method comparable in diagnostic performance to RT-qPCR.

scholarly journals Analysis of ischemic muscle in patients with peripheral artery disease using X-ray spectroscopy

2017 ◽  
Vol 220 ◽  
pp. 79-87 ◽  
Author(s):  
Ryan A. Becker ◽  
Kim Cluff ◽  
Nithyanandhi Duraisamy ◽  
George P. Casale ◽  
Iraklis I. Pipinos
Keyword(s):  
Peripheral Artery Disease ◽  
Peripheral Artery ◽  
X Ray ◽  
Artery Disease ◽  
Ischemic Muscle

scholarly journals Exercise Prior to Lower Extremity Peripheral Artery Disease Improves Endurance Capacity and Hindlimb Blood Flow by Inhibiting Muscle Inflammation

2021 ◽  
Vol 8 ◽  
Author(s):  
Maxime Pellegrin ◽  
Karima Bouzourène ◽  
Lucia Mazzolai
Keyword(s):  
Lower Extremity ◽  
Peripheral Artery Disease ◽  
Treadmill Running ◽  
Endurance Capacity ◽  
M2 Macrophage ◽  
Peripheral Artery ◽  
M1 Macrophage ◽  
Anti Inflammatory ◽  
Artery Disease ◽  
Ischemic Muscle

Lower extremity peripheral artery disease (PAD) is associated with functional decline. Physical exercise has been proven to be an effective therapeutic strategy for PAD; however the effect of exercise initiated before PAD remains unknown. Here, we investigated the preventive effects of exercise on endurance capacity, hindlimb perfusion, and on polarization profile of circulating monocytes and limb muscle macrophages. ApoE−/− mice were subjected to 5-week running wheel exercise or remained sedentary before induction of hindlimb ischemia. The two groups were thereafter kept sedentary. Exercised mice prior to PAD showed higher exhaustive treadmill running distance and time than sedentary mice. Preventive exercise also increased perfusion, arteriole density, and muscle regeneration in the ischemic hindlimb. Moreover, preventive exercise prevented ischemia-induced increased gene expression of pro-inflammatory M1 macrophages markers and cytokines in the ischemic muscle, while no changes were observed for anti-inflammatory M2 macrophage markers. Flow cytometry analysis showed that the proportion of circulating pro-inflammatory monocyte subtype decreased whereas that of anti-inflammatory monocytes increased with preventive exercise. Overall, we show that exercise initiated before PAD improves endurance performance and hindlimb perfusion in mice probably via inhibition of M1 macrophage polarization and inflammation in the ischemic muscle. Our study provides experimental evidence for a role of regular exercise in primary prevention of PAD.

A Pilot Trial of Valproic Acid in Patients with Kaposi’s Sarcoma: A Multi-Center Trial of the AIDS Malignancy Consortium.

Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 2279-2279 ◽  
Author(s):  
Mary Jo Lechowicz ◽  
Jeannette Lee ◽  
Dirk Dittmer ◽  
Susan Krown ◽  
Jonathan Said ◽  
...  
Keyword(s):  
Gene Expression ◽  
Valproic Acid ◽  
Copy Number ◽  
Kaposi's Sarcoma ◽  
Kaposi’S Sarcoma ◽  
Viral Rna ◽  
Short Term Treatment ◽  
Lytic Gene ◽  
Lytic Gene Expression ◽  
Viral Copy Number

Abstract PURPOSE: Although the use of highly active antiretroviral therapy (HAART) has led to improvements in the management of AIDS-associated Kaposi’s sarcoma (KS), KS may persist or progress despite HAART. It has been hypothesized that activation of KS-associated herpesvirus (KSHV, HHV8) lytic expression might render tumor cells susceptible to immune surveillance by cytotoxic T cells. Alternatively, it has been suggested that lytic induction could lead to tumor progression. In vitro, valproic acid (VA) and other histone deacetylase inhibitors induce KSHV lytic gene expression in primary effusion lymphoma cell lines. We investigated VA in AIDS/KS patients to assess its safety and its impact on lytic viral gene expression in tumor and viral copy number in blood. PATIENTS AND METHODS: VA was given orally to patients with AIDS and cutaneous KS on stable antiviral regimens; the dose was titrated to maintain trough concentrations between 50 and 100 mcg/mL. VA was given daily for 28 days followed by a rapid taper and patients were then followed for 6 months. Quantitative real time PCR was used to assess viral DNA in plasma and PBMC, and viral RNA in tumor specimens. Immunohistochemistry was used to assess viral antigen expression in tumor specimens. RESULTS: 18 patients were treated. 15/18 patients completed therapy; 3 patients discontinued therapy early, one secondary to grade 2 toxicity and 2 to patient preference. One patient showed a partial response and 17 showed stable disease at the completion of therapy. No patients progressed during treatment. There were no differences between KSHV copy number in plasma or PBMC before, during, or after therapy. Similarly, although serial biopsies in some patients showed an increase in lytic gene expression, these changes did not achieve statistical significance. However, in multivariate analyses, viral lytic RNA increased in tumor biopsies on day 8 as a function of VA level. There was no change in HIV viral load with VA treatment. CONCLUSION : VA was well tolerated in AIDS patients, was not associated with accelerated disease progression, but rarely induced tumor regression after short-term treatment. In patients who achieved the highest serum VA levels there was increased lytic viral RNA expression. These findings support investigation of more potent HDAC inhibitors over longer treatment courses in patients with AIDS-associated KS.

scholarly journals Effect of lyophilization on infectivity and viral load of Adenovirus

2015 ◽  
Vol 3 (1) ◽  
pp. 15-21
Author(s):  
Bimlesh Kumar Jha ◽  
Birendra Prasad Gupta ◽  
Prashanna Maharjan ◽  
Somila Kakshapati ◽  
Nabin Narayan Munankarmi
Keyword(s):  
Viral Load ◽  
Copy Number ◽  
Virus Stock ◽  
Log Reduction ◽  
Viral Copy Number ◽  
Significant Difference ◽  
Viral Copy ◽  
Viral Load Assay ◽  
Temperature And Pressure ◽  
And Storage

Freeze drying (Lyophilization) performed at temperature and pressure below the triple point is being practiced for the preservation of virus stocks for longer periods. The present study is aimed to lyophilize adenovirus strain to study its effects on infectivity and viral load. In-house adenovirus reference strain (stock virus) was propagated in Hep-2 cell line in 25cm2 cell culture flasks. In 24-well plates the serial dilutions of stock virus from 10-1 to 10-7 (100μl inoculum) was inoculated in each well with Hep-2 cells for TCID50 titer and viral DNA was extracted separately to determine viral load by Taqman Real Time PCR. Stock virus was lyophilized in 3 lots and stored at RT (25±2°C) and 4°C separately for 1, 4 and 6 months and subjected to TCID50 (for viral infectivity) and viral load assay (for total viral genome copies). Following lyophilisation and storage of adenoviral strains at RT and 4°C separately did not affect significantly on the viral stability, infectivity as well as viral copy number till 4 months. However, storage at RT for 6 months resulted in 1 log reduction in viral copy number. Thus, storage of even lyophilized virus stock would necessitate a temperature of at least 4°C for prolonged periods. The present study could successfully lyophilize adenovirus and retain its infectivity over a period of 6 months when stored at RT and 4°C. No significant difference in the infectivity or TCID50 titer was observed in the lyophilized virus as compared to the stock virus. However, the viral load was observed to increase with lyophilization of the virus over 6 months when stored at 4°C which possibly is due to the concentration of the virus on freeze-drying.Nepal Journal of Biotechnology. Dec. 2015 Vol. 3, No. 1: 15-21

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