Skip to main content. Exclusive news, data and analytics for financial market professionals Learn more about Refinitiv. CHICAGO Reuters - Carrying excess weight around the middle can impair lung function, adding to a long list of health problems associated with belly fat, French researchers said on Friday.

Abdominal obesity is already linked with diabetes, high blood pressure and heart disease as part of a cluster of health problems known collectively as metabolic syndrome.

Researchers have now shown that a large waist measurement is strongly associated with decreased lung function, regardless of other complicating factors that affect the lungs such as overall obesity and smoking.

The researchers analyzed health information about , people in France, assessing demographic background, smoking history, alcohol consumption, as well as lung function with respect to a measure of obesity known as body mass index, waist circumference and other measures of metabolic health.

Natalie Leone of the French National Institute for Health and Medical Research wrote in the American Journal of Respiratory and Critical Care Medicine.

The researchers defined abdominal obesity as having a waist circumference of greater than 35 inches for women and 40 inches for men. Several large studies have linked poor lung function with higher rates of deaths and hospitalization from heart disease, the researchers said.

While it was not clear from the study, the researchers think belly fat may impair the way the diaphragm and chest function. Fat tissue is also known to increase inflammation in the body, which may be playing a role, they said.

They were routinely tested for the absence of mycoplasma. Treatments and biological assays were carried out in duplicates on control or differentiated hMADS cells from days 4 to Total RNA was extracted using the TRI-Reagent kit Euromedex, Soufflweyersheim, France and reverse transcription RT was performed using MMLV reverse transcriptase Promega, Charbonnieres, France , as recommended by the manufacturers.

All primer sequences are described in the Supplementary section. Real-time PCR assays were run on an ABI Prism One-step real-time PCR machine Applied Biosystems, Courtaboeuf, France.

Normalization was performed using 36B4 as a reference gene. Quantification was performed using the comparative Ct method. Statistical significance was determined by t- tests BiostaTGV INSERM and Sorbonne University, PARIS, France.

HP Eschborn, Germany. Following incubation, the medium containing the inoculum was removed, the cells were washed twice, and the medium was supplemented with different specific compounds.

Uninfected cell monolayer controls were treated as the infected ones. Louis, MO, USA. Following the incubation with the virus, cells were placed in supplemented medium. Results were expressed as percentages of viable cells relative to uninfected controls.

Alterations in nuclear morphology were determined by assessment of nuclear staining using fluorescent stains and fluorescent microscopy [ 51 ]. For these experiments, hMADS adipocytes were differentiated in 2-well Lab-Tek Chamber Slides Nalge Nunc International, Naperville, IL, USA , washed with PBS pH 7.

After washing with PBS, nuclear staining was performed with Hoechst. Finally, cells were airdried and cover-slipped using Vectashield mounting medium Vector Laboratories, Burlingame, CA, USA and analyzed by fluorescent microscopy.

The number of altered nuclei were counted in the field displaying nuclear fragmentation and nuclear condensation and divided by the total number of nuclei multiplied by Observations were carried out by Lucia IMAGE 4. Lipid droplet size μm 2 was measured in SARS-CoVinfected hMADS adipocytes and untreated controls.

For this purpose, we used a drawing tablet and a morphometric program Nikon LUCIA IMAGE, Laboratory Imaging, version 4. hMADS adipocytes were examined with a Nikon Eclipse Ti-S inverted light microscope Nikon Instruments S.

A, Calenzano, Italy , and digital images were captured at ×20 with a Nikon DS-L2 camera Nikon Instruments S. A, Calenzano, Italy.

Five random fields were analyzed, at least lipid droplets were measured for each sample, and the difference between infected and non-infected cells was assessed by unpaired t -test. Similarly, the quantitative assessment of the material extruded from the hMADS was calculated using the same microscope and software and expressed as the number of vacuoles extruded from the cells on the total cell amount.

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Measurement of cell death in mammalian cells. Curr Protoc Pharmacol. Colleluori G, Graciotti L, Pesaresi M, Di Vincenzo A, Perugini J, Di Mercurio E, et al. Download references. Results from this study were made available on November 1st on bioRxiv [ 52 ]. Center for the Study of Obesity, Department of Experimental and Clinical Medicine, Marche Polytechnic University, Via Tronto 10A, Ancona, Italy.

Georgia Colleluori, Angelica Di Vincenzo, Jessica Perugini, Eleonora Di Mercurio, Cristina M. Section of Experimental Pathology, Department of Clinical and Molecular Sciences, Marche Polytechnic University, Via Tronto 10A, Ancona, Italy.

Section of Microbiology, Department of Bioscience and Public Health, Marche Polytechnic University, Via Tronto 10A, Ancona, Italy. Section of Legal Medicine, Department of Bioscience and Public Health, Marche Polytechnic University, Via Tronto 10A, Ancona, Italy. Center for Study and Research on Obesity, Department of Medical Biotechnology and Translational Medicine, University of Milan, Via Vanvitelli 32, Milan, Italy.

You can also search for this author in PubMed Google Scholar. GC, LG, MP, AG, and SC: study conceptualization. GC, LG, and SC: study coordination. MP and AT: collected autoptic samples and clinical data.

GC, MP, and ADV histological studies on autoptic samples and cell cultures. CMZ, LG, and SC: electron microscopy studies.

JP, EDM, AL, and CD: in vitro studies on hMADS. SC, PB, and SM: SARS-CoV-2 infection for the in vitro studies. GC, LG, MP, JP, EN, SM, AG, and SC: data analyses and interpretation. All authors approved the final version of the manuscript and take responsibility for its content. Correspondence to Saverio Cinti.

Reprints and permissions. Colleluori, G. et al. Int J Obes 46 , — Download citation. Received : 26 October Revised : 05 January Accepted : 12 January Published : 26 January Issue Date : May Anyone you share the following link with will be able to read this content:.

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Download PDF. Subjects Metabolic syndrome Obesity. Abstract Background Preliminary data suggested that fat embolism could explain the importance of visceral obesity as a critical determinant of coronavirus disease COVID Thoracic adipose tissue contributes to severe virus infection of the lung Article Open access 16 August SARS-CoV-2 infection induces persistent adipose tissue damage in aged golden Syrian hamsters Article Open access 01 February Association of excess visceral fat and severe illness in hospitalized COVID patients in Japan: a retrospective cohort study Article 17 January Introduction Since December , the severe acute respiratory syndrome coronavirus 2 SARS-CoV-2 , responsible for the development of coronavirus disease COVID , has spread globally, resulting in a worldwide health crisis that caused over five million deaths [ 1 ].

Results Autoptic VAT, lung and liver samples belonging to 49 subjects were collected and screened to be included in the study. Full size image. Discussion This is the first study investigating the ultrastructural features of VAT among individuals with COVID and assessing lipid distribution in lungs and liver samples by histomorphology.

Study subjects and tissue sampling Autoptic lung, liver, and VAT samples of 49 subjects were collected at the Department of Legal Medicine of the Ospedali Riuniti of Ancona between March and May In vitro studies on hMADS Ethical approval Human adipocytes progenitors -Aps- hMADS cells were isolated from adipose tissue, as surgical scraps from a surgical specimen of various surgeries of young donors, with the informed consent of the parents.

Cell differentiation hMADS cells were maintained and differentiated as previously described [ 50 ]. Gene expression analysis Total RNA was extracted using the TRI-Reagent kit Euromedex, Soufflweyersheim, France and reverse transcription RT was performed using MMLV reverse transcriptase Promega, Charbonnieres, France , as recommended by the manufacturers.

Nuclear morphology analyses Alterations in nuclear morphology were determined by assessment of nuclear staining using fluorescent stains and fluorescent microscopy [ 51 ].

References Medicine JHU. Article CAS PubMed Google Scholar Huang C, Wang Y, Li X, Ren L, Zhao J, Hu Y, et al.

Article CAS PubMed PubMed Central Google Scholar Wadman M, Couzin-Frankel J, Kaiser J, Matacic C. Article CAS PubMed Google Scholar Xu Z, Shi L, Wang Y, Zhang J, Huang L, Zhang C, et al.

Article CAS PubMed PubMed Central Google Scholar Bradley BT, Maioli H, Johnston R, Chaudhry I, Fink SL, Xu H, et al. Article CAS PubMed PubMed Central Google Scholar Schneider JL, Rowe JH, Garcia-de-Alba C, Kim CF, Sharpe AH, Haigis MC.

Visceral Fat is More Related to Impairment of Lung Function and Mechanics and Pulmonary Immune Response in Overweight and Grade I Obese Women A. Silva-Reis Santos, Brazil , M. A Rodrigues Brandao-Rangel Santos, Brazil , R. Moraes-Ferreira Santos, Brazil , T.

G Gonçalves-Alves Santos, Brazil , V. H Souza-Palmeira Santos, Brazil , H. C Aquino-Santos Santos, Brazil , C. R Frison Santos, Brazil , R. P Vieira Santos, Brazil.

Source: Virtual Congress — What's new in respiratory problems in rare and systemic diseases Session: What's new in respiratory problems in rare and systemic diseases Session type: E-poster Number: You must login to grade this presentation.

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We use cookies on our website to give you the most relevant experience by remembering your preferences and repeat visits. By clicking "Accept", you consent to the use of the cookies. P Vieira Santos, Brazil Source: Virtual Congress — What's new in respiratory problems in rare and systemic diseases Session: What's new in respiratory problems in rare and systemic diseases Session type: E-poster Number: Abstract Beyond the common comorbidities related to obesity, such as type 2 diabetes and cardiovascular diseases, impaired lung function is already known, but, whether the fat distribution sub-cutaneous, abdominal, visceral affects the lung function and pulmonary immune response are poorly known.

Thus, increased visceral fat directly influences impairment of lung function and of systemic and pulmonary immune response of obese women Rating: You must login to grade this presentation. Share or cite this content Citations should be made in the following way: A. Visceral Fat is More Related to Impairment of Lung Function and Mechanics and Pulmonary Immune Response in Overweight and Grade I Obese Women.

Member's Comments No comment yet. Related content which might interest you: Effects of Combined Physical Training on Lung Function and Mechanics and on Pulmonary Immune Response in Overweight and Obese Women Source: Virtual Congress — A multidisciplinary approach to pulmonary rehabilitation and management of chronic respiratory diseases Year: Aerobic and Weightlifting Training Improves Lung Function and Mechanics and Pulmonary and Systemic Immune Response in Overweight Women Source: Virtual Congress — Novel insights into exercise training Year: Physical Training Improves Lung Function and Mechanics and Pulmonary and Systemic Immune Response in Eutrophic Women Source: Virtual Congress — Novel insights into exercise training Year: Resistance Training Alone Improves Lung Inflammation and Respiratory Muscle Strenght but not Lung Function and Mechanics in Mid-Age Women Source: Virtual Congress — Novel insights into exercise training Year: The Dietary Fiber Intake and its Relationship of Lung Function and Inflammatory Markers in Korean Adults Source: International Congress — Epidemiology of airway diseases in primary care Year: Respiratory Symptoms and Lung Function Depend on Smoking Status and Enviromental Conditions Source: International Congress — Smoking, apnoea, infection, allergens and other factors Year: A Light microscopy LM : hyaline membranes lining alveolar surfaces arrows at low magnification.

Lipid-rich macrophages free in the alveolar space red arrows and inside hyaline membranes red arrow. D LM: enlargement of the squared area in C. Arrows indicate lipid vacuoles.

F TEM: free lipid droplets lining the alveolar surface composed by pneumocytes type II PT2 with classic surfactant granules arrow. Lastly, since the embolic material from abdominal visceral tissues should necessarily pass through the liver parenchyma to reach the lung, we exploited the ORO staining technique to study liver samples belonging to 9 individuals with COVID and 8 control subjects.

Liver autoptic samples showed focal, macrovesicular steatosis with lipid droplets of very variable size Supplementary Fig. In particular, signs consistent with fat embolism, i. This is the first study investigating the ultrastructural features of VAT among individuals with COVID and assessing lipid distribution in lungs and liver samples by histomorphology.

Our data support the presence of higher local VAT inflammation and higher prevalence of fat embolism and lipidic HM formations in the lungs of subjects dead due to COVID compared to control individuals dead for different reasons.

In addition, our data support SARS-CoV-2 ability to infect human adipocytes in vitro. Considering the strong association between COVIDrelated complications and obesity, especially with visceral adipose content excess [ 10 , 11 , 13 , 14 , 15 , 16 ], the comprehension of the biological phenomenon at the basis of such association holds critical clinical implications in the era of the COVID pandemic.

Our study provides the first evidence of higher local VAT inflammation among subjects with COVID, independently of obesity status and support an exacerbation of obesity-related inflammation by SARS-CoV-2 infection, a novel finding consistent with studies reporting higher systemic inflammation among infected patients [ 18 ].

Adipocyte inflammation is associated with cell stress, death, and lipid release in the extracellular space [ 19 , 20 , 25 , 26 ]. We hence studied adipocyte features by TEM and revealed the presence of the typical signs of cellular stress, together with prominent features of lipids spill-over from suffering adipocytes.

Of note, these data are supported by a recent work showing an increased number of autoimmune antibodies against the malondialdehyde and the adipocyte-derived protein antigen markers of lipid peroxidation and adipocytes death, respectively [ 35 ] among subjects with COVID and obesity as compared to individuals suffering from each condition independently [ 36 ].

In addition, hyperglycemia among subjects with COVID was demonstrated to be strongly associated with insulin resistance and low plasma adiponectin levels [ 29 ]. The authors from the same study also demonstrated that SARS-CoV-2 could infect hamster adipose tissue, leading to reduced adiponectin production and speculated that SARS-CoV-2 infection might result in adipocyte dysfunction driving insulin resistance.

Importantly, we detected lipids in the extracellular space, inside endothelial cells, inside the capillary lumen, and extruding from endothelial cells into the capillary lumen, all features indicative of fat embolism. Although virus-like structures were evidenced by TEM in the same VAT depots, the lack of SARS-CoV-2 detection by qPCR did not allow us to conclude that such inflammation, cellular stress, and death were all related to the presence of the virus.

It is, in fact, possible that the described VAT features were secondary to the systemic inflammation induced by COVID or due to the presence of different viruses within the depot. On the other side, we demonstrated that SARS-CoV-2 could infect human adipocytes even though neither adipocytes nor adipocyte progenitors gathered all of the known molecular requirements for the virus entry e.

This set of data is in part consistent with other findings and suggests that additional, not yet characterized, receptors and proteases may be exploited for this purpose [ 16 , 37 ].

Puray-Chavez et al. in fact indicated that human H lung adenocarcinoma cells are permissive to SARS-CoV-2 infection despite complete ACE2 absence and that virus entry is dependent on heparan sulfate in this cell line [ 37 ]. Importantly, despite being the first SARS-CoV-2 targets, olfactory and respiratory epithelial cells express low ACE2 protein levels [ 38 ].

For these reasons, additional co-factors facilitating the virus-host cell interaction e. In our study, BASIGIN receptor and FURIN protease were highly expressed in differentiated hMADS and could be exploited for SARS-CoV-2 infection.

However, it should be noted that, although FURIN critical role in mediating SARS-CoV-2 infection is widely accepted and seem to be of relevance in patients with type 2 diabetes where the protease is highly expressed [ 41 ], the role of BASIGIN has been recently questioned [ 42 ].

Given the widespread presence of lipid droplets in the capillary lumen of VAT and our preliminary data [ 23 ], we studied lipid distribution in lung and liver samples and confirmed the presence of fat embolism.

Fat embolism in the lungs was not exclusive to, but more prevalent among subjects with COVID; it was in fact also detected among subjects with obesity independently of SARS-CoV-2 infection. These data are not surprising given that adipocyte death and release of lipids are both phenomena occurring in obesity [ 19 , 25 , 26 ].

This finding provides the first evidence pointing out fat embolism as a complication of obesity and obesity plus type 2 diabetes , determined by adipocyte death and possibly exacerbated by the COVIDinduced inflammatory status. HM were present in all patients with COVID and in only one control who died for pneumonia, a finding consistent with other reports describing HM presence in this latter disease [ 7 ].

Our histomorphologic assessment revealed several aspects indicative of a direct role of embolic fat in HM formation. Consistently, the presence of lung HM of lipidic nature in the lungs was associated with VAT inflammation. Our findings on intestinal and liver fat embolism strongly support the embolic nature of lipid droplets in the lungs.

As the portal system drains venous blood from most abdominal fat depots to the liver, the embolic fat originated in the VAT necessarily pass through the liver to reach other organs. The unequivocal presence of lipid droplets into sinusoids and liver veins supports the fat embolic production by abdominal fat.

In summary, in our case series, although fat embolism may be present in obesity and type 2 diabetes independently of COVID, the embolic lipidic material could contribute to the formation of HM only in the case of COVIDrelated pneumonia.

This novel finding holds critical clinical implications and deserves further investigation. Furthermore, these data provide insights into HM nature, as their formation process has not been characterized yet [ 44 ].

Additional studies investigating the HM nature of non-COVIDrelated pneumonia are required to detail such histopathological features. Collectively our data reveal higher local VAT inflammation in subjects with COVID and SARS-CoV-2 ability to infect human adipocytes.

In addition, we provide the first evidence that supports the fat embolism as a complication of obesity, likely determined by adipocyte death and exacerbated by the COVIDinduced inflammatory status.

Consistently, fat embolism displays similar signs and symptoms as observed in COVID, in line with a recently published case report [ 45 ].

When fat embolism and COVID are suspected, differential diagnosis is critical for proper patient care. Based on our findings, the assessment of fat embolism symptoms is mandatory in the context of the COVID pandemic, especially among patients with pulmonary symptoms, obesity, and high waist circumference, last two of which are recognized as signs of high visceral adipose accumulation.

Such complex clinical status should be therefore adequately assessed and properly addressed. Our data hold critical clinical implications in the context of obesity and COVID pandemics and need to be confirmed by additional studies with larger sample size.

Our study did not entail any physical risk for the subjects. In Italy, the evaluation of non-pharmacological observational studies is not governed by the same normative references provided for the evaluation of clinical trials and observational studies concerning drugs. Furthermore, as reported in the above report [ 46 ] in the section dedicated to our type of study in conditions of pandemic and therefore of high risk for the communities, some administrative steps may be abolished.

Therefore, our Institutional Review Board does not require ethical approval for studies conducted on autoptic specimens and not collecting personal or sensitive data.

Autoptic lung, liver, and VAT samples of 49 subjects were collected at the Department of Legal Medicine of the Ospedali Riuniti of Ancona between March and May Twenty-four subjects were affected by COVID, while the remaining 25 were not and died for different reasons.

SARS-CoV-2 infection was assessed in all subjects by RT-PCR tests on a nasopharyngeal swab. Among the studied subjects, 15 had documented respiratory conditions, i.

VAT was sampled from the omentum and mesentery region. Lungs were extensively sampled across central and peripheral regions of each lobe bilaterally. A median of seven tissue blocks range five to nine were taken from each lung.

Liver samples were collected from the right and left lobes. Samples were sliced into different pieces to be studied by LM and transmission electron microscopy TEM.

A comprehensive methodological description for such methodologies has been described elsewhere [ 47 ]. Samples were then embedded in paraffin to be studied by LM and to perform immunohistochemistry and morphometric analyses. A comprehensive description of the protocol has been described elsewhere [ 47 ].

To study SARS-CoV-2 presence in VAT, we used the SARS-CoV-2 nucleocapsid Invitrogen MA and spike protein Sino Biological T62 antibodies at different dilutions. The same antibodies were used to detect the virus on infected VeroE6 at dilution: for nucleocapsid protein and for the spike protein.

Negative control in which primary antibody was omitted were always included in each set of reactions to assess antibody specificity.

Tissue sections were observed with a Nikon Eclipse E light microscope. For morphometric purposes, for each paraffin section, ten digital images were acquired at ×20 magnification with a Nikon DXM camera.

Tissue slices were then counterstained with hematoxylin and covered with a cover-slip using Vectashield mounting medium Vector Laboratories. The day before infection, a confluent monolayer was trypsinized, and 1. Confluent monolayers were infected with SARS-CoV-2 isolates, accession no.

MT [ 49 ] at a multiplicity of infection of 3. Uninfected cell monolayer controls were treated as infected ones. Aliquots of infected supernatants, collected as above, were analyzed using RT-qPCR assay as described elsewhere [ 49 ].

Human adipocytes progenitors -Aps- hMADS cells were isolated from adipose tissue, as surgical scraps from a surgical specimen of various surgeries of young donors, with the informed consent of the parents. All methods were approved and performed following the guidelines and regulations of the Centre Hospitalier Universitaire de Nice Review Board.

hMADS cells were maintained and differentiated as previously described [ 50 ]. They will be further referred to as hMADS adipocytes.

They were routinely tested for the absence of mycoplasma. Treatments and biological assays were carried out in duplicates on control or differentiated hMADS cells from days 4 to Total RNA was extracted using the TRI-Reagent kit Euromedex, Soufflweyersheim, France and reverse transcription RT was performed using MMLV reverse transcriptase Promega, Charbonnieres, France , as recommended by the manufacturers.

All primer sequences are described in the Supplementary section. Real-time PCR assays were run on an ABI Prism One-step real-time PCR machine Applied Biosystems, Courtaboeuf, France.

Normalization was performed using 36B4 as a reference gene. Quantification was performed using the comparative Ct method.

Statistical significance was determined by t- tests BiostaTGV INSERM and Sorbonne University, PARIS, France. HP Eschborn, Germany. Following incubation, the medium containing the inoculum was removed, the cells were washed twice, and the medium was supplemented with different specific compounds.

Uninfected cell monolayer controls were treated as the infected ones. Louis, MO, USA. Following the incubation with the virus, cells were placed in supplemented medium. Results were expressed as percentages of viable cells relative to uninfected controls.

Alterations in nuclear morphology were determined by assessment of nuclear staining using fluorescent stains and fluorescent microscopy [ 51 ]. For these experiments, hMADS adipocytes were differentiated in 2-well Lab-Tek Chamber Slides Nalge Nunc International, Naperville, IL, USA , washed with PBS pH 7.

After washing with PBS, nuclear staining was performed with Hoechst. Finally, cells were airdried and cover-slipped using Vectashield mounting medium Vector Laboratories, Burlingame, CA, USA and analyzed by fluorescent microscopy. The number of altered nuclei were counted in the field displaying nuclear fragmentation and nuclear condensation and divided by the total number of nuclei multiplied by Observations were carried out by Lucia IMAGE 4.

Lipid droplet size μm 2 was measured in SARS-CoVinfected hMADS adipocytes and untreated controls. For this purpose, we used a drawing tablet and a morphometric program Nikon LUCIA IMAGE, Laboratory Imaging, version 4. hMADS adipocytes were examined with a Nikon Eclipse Ti-S inverted light microscope Nikon Instruments S.

A, Calenzano, Italy , and digital images were captured at ×20 with a Nikon DS-L2 camera Nikon Instruments S. A, Calenzano, Italy. Five random fields were analyzed, at least lipid droplets were measured for each sample, and the difference between infected and non-infected cells was assessed by unpaired t -test.

Similarly, the quantitative assessment of the material extruded from the hMADS was calculated using the same microscope and software and expressed as the number of vacuoles extruded from the cells on the total cell amount.

Statistical analyses were performed with Prism 6. Medicine JHU. Coronavirus resource center. Mahendra M, Nuchin A, Kumar R, Shreedhar S, Mahesh PA. Predictors of mortality in patients with severe COVID pneumonia—a retrospective study. Adv Respir Med. Article CAS PubMed Google Scholar.

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