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Coexistence of antiphospholipid antibodies and cephalalgia

Islam MA, Alam F, Gan SH, Cavestro C, Wong KK

Cephalalgia, 2018 03;38(3):568-580.
PMID: 28952322 DOI: 10.1177/0333102417694881

Background The occurrence of antiphospholipid antibodies (aPLs) and headache comorbidity in the presence or absence of underlying autoimmune diseases remains unclear. Aim The aim of this review was to summarize the relationship between headache and aPLs based on evidences from cohort studies and case reports, in addition to examining the treatment strategies that resolved headache in aPLs-positive individuals.
Methods A comprehensive literature search was conducted through PubMed, ISI Web of Science and Google Scholar. A total of 559 articles were screened and the appropriate articles were selected based on quality and level of evidence.
Results Cohort studies (n = 27) from Europe, North America and Asia demonstrated comorbidity of aPLs and headache in antiphospholipid syndrome, systemic lupus erythematosus (SLE) and neuropsychiatric SLE patients. Significantly higher association between migraine and aPLs was observed (n = 170/779; p

Matched MeSH terms: Antibodies, Antiphospholipid/blood*
Integration of molecular dynamics simulation and hotspot residues grafting for de novo scFv design against Salmonella Typhi TolC protein

Leong SW, Lim TS, Ismail A, Choong YS

J. Mol. Recognit., 2018 05;31(5):e2695.
PMID: 29230887 DOI: 10.1002/jmr.2695

With the development of de novo binders for protein targets from non-related scaffolds, many possibilities for therapeutics and diagnostics have been created. In this study, we described the use of de novo design approach to create single-chain fragment variable (scFv) for Salmonella enterica subspecies enterica serovar Typhi TolC protein. Typhoid fever is a global health concern in developing and underdeveloped countries. Rapid typhoid diagnostics will improve disease management and therapy. In this work, molecular dynamics simulation was first performed on a homology model of TolC protein in POPE membrane bilayer to obtain the central structure that was subsequently used as the target for scFv design. Potential hotspot residues capable of anchoring the binders to the target were identified by docking "disembodied" amino acid residues against TolC surface. Next, scFv scaffolds were selected from Protein Data Bank to harbor the computed hotspot residues. The hotspot residues were then incorporated into the scFv scaffold complementarity determining regions. The designs recapitulated binding energy, shape complementarity, and interface surface area of natural protein-antibody interfaces. This approach has yielded 5 designs with high binding affinity against TolC that may be beneficial for the future development of antigen-based detection agents for typhoid diagnostics.

Matched MeSH terms: Single-Chain Antibodies/chemistry*
Human neonatal Fc receptor as a new potential antibody binding protein for antibody immobilization

Ng WK, Lim TS, Lai NS

Biotechnol Appl Biochem, 2018 Jul;65(4):547-553.
PMID: 29280199 DOI: 10.1002/bab.1636

A critical challenge in producing an antibody-based assay with the highest reproducibility and sensitivity is the strategy to immobilize antibodies to solid phase. To date, numerous methods of antibody immobilization were reported but each was subjected to its advantages and limitations. The current study proposes a new potential antibody binding protein, the human neonatal fragment crystallizable (Fc) receptor. This protein has shown its high affinity to the Fc of antibody either in vivo or in vitro. Human neonatal Fc receptor is a heterodimer constructed by p51 α-heavy chain and β2-microglobulin light chain; however, the binding sites toward the antibody are located in the p51 α-heavy chain. Hence, vector cloning and recombinant protein expression were carried out to express the p51 α-heavy chain of the human neonatal Fc receptor (hFcRn-α). The recombinant protein expressed, hFcRn-α, was adopted to pin rabbit IgG against hepatitis B virus surface antigen to a solid phase. A sandwich enzyme-linked immunosorbent assay was further developed to evaluate the efficiency of hFcRn-α-directed immobilization in antigen detection. The result was compared with the conventional physical adsorption method. The findings demonstrated that human neonatal Fc receptor was efficient in pinning antibodies and generating higher signals compared with the physical adsorption of antibody.

Matched MeSH terms: Antibodies, Immobilized/immunology*
Single-chain Fv Antibodies for Targeting Neurodegenerative Diseases

Chia KY, Ng KY, Koh RY, Chye SM

CNS Neurol Disord Drug Targets, 2018;17(9):671-679.
PMID: 29546836 DOI: 10.2174/1871527317666180315161626

BACKGROUND & OBJECTIVE: Protein misfolding and aggregation have been considered the common pathological hallmarks for a number of neurodegenerative diseases, including Alzheimer's disease (AD), Parkinson's disease (PD) and Huntington's disease (HD). These abnormal proteins aggregates damage mitochondria and induce oxidative stress, resulting in neuronal cell death. Prolonged neuronal damage activates microglia and astrocytes, development of inflammation reaction and further promotes neurodegeneration. Thus, elimination of abnormal protein aggregates without eliciting any adverse effects are the main treatment strategies. To overcome this, recent studies have deployed single- chain fragment variable antibodies (scFvs) to target the pathological protein aggregates, such as amyloid-beta (Aβ) peptides, α-synuclein (α-syn) and Huntingtin (Htt). To date scFv has been effective at inhibiting abnormal protein aggregates formation in both in vitro and in vivo model system of AD, PD and HD.
CONCLUSION: Currently active research is still ongoing to improve the scFv gene delivery technology, to further enhance brain penetration, intracellular stability, solubility and efficacy of scFv intrabody.

Matched MeSH terms: Single-Chain Antibodies/therapeutic use*
Fulltext Cross-Neutralizing CRF01_AE-Infected Plasma from Malaysia Targets CD4-Binding Site of Human Immunodeficiency Virus Type-1 Envelope Glycoprotein

Ng QR, Tee KK, Binley JM, Tong T

AIDS Res Hum Retroviruses, 2022 Feb;38(2):162-172.
PMID: 34006141 DOI: 10.1089/AID.2020.0299

Human immunodeficiency virus type-1 (HIV-1) antigenic variation poses a great challenge for vaccine immunogen design to elicit broadly neutralizing antibodies (bNAbs). Over the last 10-15 years, great progress has been made to understand the conserved sites of sensitivity on HIV envelope glycoprotein spikes targeted by bNAbs. Plasma neutralization mapping and monoclonal antibody isolation efforts have revealed five major conserved epitope clusters. Most of this work has focused on subtype B and C-infected Caucasian or African donors. It is not clear if the same epitopes and epitope rank order preferences are also true in donors infected with different HIV-1 subtypes and with different racial backgrounds. To investigate this point, in this study we report the first attempt to profile the bNAb specificities of CRF01_AE-infected Malaysian plasmas. We first measured neutralization titers of 21 plasmas against a subtype A, B, and AE pseudovirus panel. This revealed that 14% (3 of 21) plasmas had cross-clade breadth. Focusing on the cross-neutralizing plasma P9, we used AE and JR-FL mutant pseudoviruses, gp120 monomer interference, and native polyacrylamide gel electrophoresis to better understand the neutralization specificity. P9 demonstrates CD4-binding-site specificity with trimer dependence and D368 independence.

Matched MeSH terms: HIV Antibodies; Antibodies, Neutralizing
Anti-Tumor Necrosis Factor for Supplementary Management in Severe Asthma: A Systematic Review and Meta-analysis

Ahmad S, Mohd Noor N, Engku Nur Syafirah EAR, Irekeola AA, Shueb RH, Chan YY, et al.

J Interferon Cytokine Res, 2023 Feb;43(2):77-85.
PMID: 36795972 DOI: 10.1089/jir.2022.0211

Tumor-necrosis factor (TNF) is recognized as a therapeutic target in inflammatory diseases, including asthma. In severe forms of asthma, biologics such as anti-TNF are rendered to be investigated as therapeutic options in severe asthma. Hence, this work is done to assess the efficacy and safety of anti-TNF as a supplementary therapy for patients with severe asthma. A systematic search of 3 databases (Cochrane Central Register of Controlled Trials, MEDLINE, ClinicalTrials.gov) was performed to identify for published and unpublished randomized controlled trials comparing anti-TNF (etanercept, adalimumab, infliximab, certolizumab pegol, golimumab) with placebo in patients diagnosed with persistent or severe asthma. Random-effects model was used to estimate risk ratios and mean differences (MDs) with confidence intervals (95% CIs). PROSPERO registration number is CRD42020172006. Four trials with 489 randomized patients were included. Comparison between etanercept and placebo involved 3 trials while comparison between golimumab and placebo involved 1 trial. Etanercept produced a small but significant impairment in forced expiratory flow in 1 second (MD 0.33, 95% CI 0.09-0.57, I2 statistic = 0%, P = 0.008) and a modest improvement of asthma control using the Asthma Control Questionnaire. However, using the Asthma Quality of Life Questionnaire, the patients exhibit an impaired quality of life with etanercept. Treatment with etanercept showed a reduced injection site reaction and gastroenteritis compared with placebo. Although treatment with anti-TNF is shown to improve asthma control, severe asthma patients did not benefit from this therapy as there is limited evidence for improvement in lung function and reduction of asthma exacerbation. Hence, it is unlikely to prescribe anti-TNF in adults with severe asthma.

Matched MeSH terms: Antibodies, Monoclonal, Humanized/therapeutic use
Fulltext Seroprevalence of anti-HCV in an urban child population: a preliminary study from Kuala Lumpur

Lee WS, Ng KP

Singapore Med J, 2001 Mar;42(3):100-1.
PMID: 11405558

A pilot study to determine the seroprevalence of anti-HCV among children from Kuala Lumpur, Malaysia, was conducted using microparticle enzyme immunoassay. Serum samples were obtained randomly from children, aged between one to 16 years of age, admitted to the paediatric unit of University of Malaya Medical Centre, Kuala Lumpur for various medical reasons. Of the 179 samples assayed, only one was positive, giving the prevalence rate of 0.6%. It is reasonable to conclude that the seroprevalence of anti-HCV among children from Kuala Lumpur is low, less than 1%.

Matched MeSH terms: Hepatitis C Antibodies/blood*
Fulltext The increasing prevalence of endemic typhus in Kuala Lumpur and an evaluation of a diagnostic ELISA dot test for the detection of antibodies to Rickettsia typhi

Sekhar WY, Devi S

Singapore Med J, 2000 May;41(5):226-31.
PMID: 11063173

A seroepidemiology study was done in response to the recent increase of Endemic Typhus cases diagnosed at University Hospital. The serosurvey was based on doctors' request for the Weil Felix (WF) or the Indirect Immunoperoxidase (IIP) test in Pyrexia of Unknown Origin (PUO) patients for the years 1991 to 1997. Over the 7 years, we found that the incidence of Endemic typhus is increasing with gender (male:female = 2:1), age (20-40 years) and race distribution (Indians > Malay > Chinese) that reflects socioeconomic circumstances. A commercially available ELISA dot assay [INDX (E2R3) Dip-S-Ticks], for the detection of antibodies against R. typhi was compared with the indirect immunoperoxidase test (IIP). The ELISA assay was done against 219 IIP tested sera. The Dip-S-Ticks was found to be comparable to the IIP with a sensitivity of 91.7% and specificity of 92.8% at cut-off titres of > 1:80 IIP.

Matched MeSH terms: Antibodies, Bacterial/analysis*
A large exposure to Brucella melitensis in a diagnostic laboratory

Sam IC, Karunakaran R, Kamarulzaman A, Ponnampalavanar S, Syed Omar SF, Ng KP, et al.

J Hosp Infect, 2012 Apr;80(4):321-5.
PMID: 22237130 DOI: 10.1016/j.jhin.2011.12.004

Brucella species are easily transmitted by aerosols and can be acquired in the laboratory.

Matched MeSH terms: Antibodies, Bacterial/blood
Fulltext Outcomes of tocilizumab therapy in severe or critical COVID-19 patients: A retrospective cohort, single-centre study

Abdelnaby H, Aboelhassan W, Al-Jarallah M, Rajan R, Dashti R, Zhanna KD, et al.

Trop Med Int Health, 2021 Dec;26(12):1689-1699.
PMID: 34601803 DOI: 10.1111/tmi.13685

OBJECTIVES: To assess the effectiveness and safety of tocilizumab, a humanised anti-interleukin-6 receptor antibody, in the treatment of critical or severe coronavirus disease 2019 (COVID-19) patients.
METHODS: This was a retrospective cohort study of severe or critical COVID-19 patients (≥18 years) admitted to one hospital in Kuwait. Fifty-one patients received intravenous tocilizumab, while 78 patients received the standard of care at the same hospital. Both groups were compared for clinical improvement and in-hospital mortality.
RESULTS: The tocilizumab (TCZ) group had a significantly lower 28-day in-hospital mortality rate than the standard-of care-group (21.6% vs. 42.3% respectively; p = 0.015). Fifty-five per cent of patients in the TCZ group clinically improved vs. 11.5% in the standard-of-care group (p

Matched MeSH terms: Antibodies, Monoclonal, Humanized/therapeutic use*
Dot immunogold filtration assay (DIGFA) for the rapid detection of specific antibodies against the rat lungworm Angiostrongylus cantonensis (Nematoda: Metastrongyloidea) using purified 31-kDa antigen

Eamsobhana P, Gan XX, Ma A, Wang Y, Wanachiwanawin D, Yong HS

J Helminthol, 2014 Dec;88(4):396-401.
PMID: 23710755 DOI: 10.1017/S0022149X13000321

A rapid dot immunogold filtration assay (DIGFA) was adopted for specific immunodiagnosis of human cerebral angiostrongyliasis, using purified 31-kDa glycoprotein specific to Angiostrongylus cantonensis as diagnostic antigen and protein A colloidal gold conjugate as antigen-antibody detector. A total of 59 serum samples were assayed - 11 samples from clinically diagnosed patients with detectable A. cantonensis-specific antibody in immunoblotting; 23 samples from patients with other related parasitic diseases, i.e. gnathostomiasis (n= 8), cysticercosis (n= 5), toxocariasis (n= 2), filariasis (n= 4), paragonimiasis (n= 2) and malaria (n= 2); and 25 samples from normal healthy subjects. The sensitivity and specificity of DIGFA to detect anti-A. cantonensis specific antibodies in serologically confirmed angiostrongyliasis cases, were both 100%. No positive DIGFA was observed in cases with other parasitic diseases, and the healthy control subjects. The 3-min DIGFA is as sensitive and specific as the 3-h immunoblot test in angiostrongyliasis confirmed cases that revealed a 31-kDa reactive band. The gold-based DIGFA is more rapid and easier to perform than the traditional enzyme-linked immunosorbent assay (ELISA). The test utilizing purified A. cantonensis antigen is reliable and reproducible for specific immunodiagnosis of human infection with A. cantonensis - thus can be applied as an additional routine test for clinical diagnostic support. Large-scale sero-epidemiological studies in endemic communities in north-east Thailand are under way to evaluate its usefulness under field conditions.

Matched MeSH terms: Antibodies, Helminth/immunology*
Letter: Posttransfusion hepatitis in Kuala Lumpur

Kamath S, Lopez CG

Med J Aust, 1973 Nov 3;2(18):867-8.
PMID: 4782397
Matched MeSH terms: Hepatitis B Antibodies/analysis
Fulltext Evaluation of Elecsys Syphilis Assay for Routine and Blood Screening and Detection of Early Infection

Kremastinou J, Polymerou V, Lavranos D, Aranda Arrufat A, Harwood J, Martínez Lorenzo MJ, et al.

J Clin Microbiol, 2016 09;54(9):2330-6.
PMID: 27358468 DOI: 10.1128/JCM.02544-15

Treponema pallidum infections can have severe complications if not diagnosed and treated at an early stage. Screening and diagnosis of syphilis require assays with high specificity and sensitivity. The Elecsys Syphilis assay is an automated treponemal immunoassay for the detection of antibodies against T. pallidum The performance of this assay was investigated previously in a multicenter study. The current study expands on that evaluation in a variety of diagnostic settings and patient populations, at seven independent laboratories. The samples included routine diagnostic samples, blood donation samples, samples from patients with confirmed HIV infections, samples from living organ or bone marrow donors, and banked samples, including samples previously confirmed as syphilis positive. This study also investigated the seroconversion sensitivity of the assay. With a total of 1,965 syphilis-negative routine diagnostic samples and 5,792 syphilis-negative samples collected from blood donations, the Elecsys Syphilis assay had specificity values of 99.85% and 99.86%, respectively. With 333 samples previously identified as syphilis positive, the sensitivity was 100% regardless of disease stage. The assay also showed 100% sensitivity and specificity with samples from 69 patients coinfected with HIV. The Elecsys Syphilis assay detected infection in the same bleed or earlier, compared with comparator assays, in a set of sequential samples from a patient with primary syphilis. In archived serial blood samples collected from 14 patients with direct diagnoses of primary syphilis, the Elecsys Syphilis assay detected T. pallidum antibodies for 3 patients for whom antibodies were not detected with the Architect Syphilis TP assay, indicating a trend for earlier detection of infection, which may have the potential to shorten the time between infection and reactive screening test results.

Matched MeSH terms: Antibodies, Bacterial/blood
Fulltext Formaldehyde-inactivated whole-virus vaccine protects a murine model of enterovirus 71 encephalomyelitis against disease

Ong KC, Devi S, Cardosa MJ, Wong KT

J Virol, 2010 Jan;84(1):661-5.
PMID: 19864378 DOI: 10.1128/JVI.00999-09

Enterovirus 71 (EV71) causes childhood hand, foot, and mouth disease and neurological complications, and no vaccines or therapeutic drugs are currently available. Formaldehyde-inactivated whole-virus vaccines derived from EV71 clinical isolates and a mouse-adapted virus (MAV) were tested in a mouse model of EV71 encephalomyelitis. After only two immunizations, given to mice at 1 and 7 days of age, the MAV vaccine protected mice at 14 days of age from disease. Tissues from immunized mice were negative for virus by viral culture, reverse transcriptase PCR, immunohistochemistry analysis, and in situ hybridization. Cross-neutralizing EV71 antibodies to strains with genotypes B3, B4, and C1 to C5 generated in immunized adult mice were able to passively protect 14-day-old mice from disease.

Matched MeSH terms: Antibodies, Neutralizing/immunology
Secukinumab therapy for paediatric patients with various phenotypes of congenital ichthyosis

Albela H, Ting IPL, Lee TS, Ooi D, Leong KF

Clin Exp Dermatol, 2024 Jul 19;49(8):920-922.
PMID: 38470562 DOI: 10.1093/ced/llae082
Matched MeSH terms: Antibodies, Monoclonal/therapeutic use
Newcastle disease vaccination in Malaysia: application of oil emulsion vaccine

Ernawati R, Ibrahim AL

Vet Rec, 1984 Oct 06;115(14):352-4.
PMID: 6495601

An experimental oil emulsion Newcastle disease vaccine was evaluated for its efficacy in broiler chickens. A group of chickens vaccinated at one day old with a live lentogenic Newcastle disease vaccine and subsequently revaccinated at three and eight weeks old with the experimental oil emulsion vaccine showed satisfactory haemagglutination inhibition antibody response which persisted for 18 weeks. Between 90 and 100 per cent of the vaccinated chickens were protected when challenged with the velogenic viscerotropic Newcastle disease virus. Although the vaccinated chickens were protected against clinical disease, virus could be isolated from a number of birds. By day 10 to 12 after challenge all the chickens were free from Newcastle disease infection.

Matched MeSH terms: Antibodies, Viral/analysis
Fulltext Don't close the book on tocilizumab for the treatment of severe COVID-19 pneumonia-the jury is still out: The Kuwait experience

Al-Shamali Y, M Ali Y, Al-Shamali RA, Al-Melahi M, Al-Shammari FR, Alsaber A, et al.

PLoS One, 2021;16(8):e0254379.
PMID: 34428204 DOI: 10.1371/journal.pone.0254379

PURPOSE: This cross-sectional observational study aims to report preliminary data from the first experience using tocilizumab for patients with severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) infection in three of Kuwait's largest public hospitals City.
PATIENTS AND METHODS: This chart review study examined the benefits of tocilizumab treatment among 127 patients diagnosed with severe coronavirus disease of 2019 (COVID-19) pneumonia.
RESULTS: 90 of 127 patients (71%) survived. Mortality was highest in the elderly with multiple medical conditions.
CONCLUSION: Despite the small sample size and retrospective nature of the work, our findings are consistent with recent studies suggesting tocilizumab administration in patients presenting with severe COVID pneumonia with associated hyperinflammatory features conferred mortality benefit.

Matched MeSH terms: Antibodies, Monoclonal, Humanized/therapeutic use*
Fulltext Prevalence of Toxoplasma gondii Antibodies in Stray Dogs from Various Locations in West and East Malaysia

Watanabe M, Sadiq MB, Mulop NIA, Mohammed K, Rani PAM, Fong LS, et al.

Korean J Parasitol, 2020 Oct;58(5):487-492.
PMID: 33202500 DOI: 10.3347/kjp.2020.58.5.487

Toxoplasmosis is caused by an obligate intracellular protozoan parasite; Toxoplasma gondii, which is one of the most important zoonotic parasite worldwide. In dogs, the sexual reproductive cycle of T. gondii is lacking, and the animals are not widely consumed as food, but they are vital in the mechanical transmission of the parasite. However, there is no present data on the exposure of stray dogs to T. gondii in Malaysia. The objective of this serological survey was to determine the prevalence of T. gondii antibodies (IgG) and associated factors in stray dogs in East and West Malaysia. Antibodies to T. gondii were determined in serum samples from 222 stray dogs from 6 different states in East and West Malaysia (Peninsular Malaysia) using an Indirect ELISA. The seroprevalence for T. gondii was 23.4% (Confidence interval: CI 17.8-29.2%). Stray dogs from Selangor and Kuala Lumpur had the highest seroprevalence (32.4%; CI 13.2-45.5%) and lowest in those from Penang and Kedah (12.5%; CI 1.3-23.5%). Gender and breed were not associated with T. gondii seropositivity. However, adult dogs were more likely to be seropositive for T. gondii (OR=2.89; CI 1.1-7.7) compared with younger dogs. These results revealed that T. gondii is prevalent in stray dogs in the studied areas in Malaysia, and indicative of the level of environmental contamination of this parasite especially in urban areas.

Matched MeSH terms: Antibodies, Protozoan/analysis*
Fulltext Immune-mediated necrotizing myopathy (NAM) related to SARS-Cov-2 infection: a case report

Ibrahim A, Ghazali WSW, Misyail A, Najwa L, Khan AH, Amir WM, et al.

BMC Neurol, 2023 Mar 22;23(1):117.
PMID: 36949469 DOI: 10.1186/s12883-023-03170-1

BACKGROUND: There is a growing body of evidence that severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) or COVID-19 infection is associated with the development of autoimmune diseases. A recent systematic review reported that the new-onset autoimmune disorders during or after COVID-19 infection included inflammatory myopathies such as immune-mediated necrotizing myopathies.
CASE PRESENTATION: We described a 60-year-old man diagnosed with COVID-19 infection and later presented with a two-week history of myalgia, progressive limb weakness, and dysphagia. He had a Creatinine Kinase (CK) level of more than 10,000 U/L, was strongly positive for anti-signal recognition particle (SRP) and anti-Ro52 antibody, and a muscle biopsy revealed a paucity-inflammation necrotizing myopathy with randomly distributed necrotic fibers, which was consistent with necrotizing autoimmune myositis (NAM). He responded well clinically and biochemically to intravenous immunoglobulin, steroids and immunosuppressant and he was able to resume to his baseline.
CONCLUSION: SARS-CoV-2 may be associated with late-onset necrotizing myositis, mimicking autoimmune inflammatory myositis.

Matched MeSH terms: Antibodies, Antinuclear/blood
Fulltext Recent advances in application of biosensors in tissue engineering

Hasan A, Nurunnabi M, Morshed M, Paul A, Polini A, Kuila T, et al.

Biomed Res Int, 2014;2014:307519.
PMID: 25165697 DOI: 10.1155/2014/307519

Biosensors research is a fast growing field in which tens of thousands of papers have been published over the years, and the industry is now worth billions of dollars. The biosensor products have found their applications in numerous industries including food and beverages, agricultural, environmental, medical diagnostics, and pharmaceutical industries and many more. Even though numerous biosensors have been developed for detection of proteins, peptides, enzymes, and numerous other biomolecules for diverse applications, their applications in tissue engineering have remained limited. In recent years, there has been a growing interest in application of novel biosensors in cell culture and tissue engineering, for example, real-time detection of small molecules such as glucose, lactose, and H2O2 as well as serum proteins of large molecular size, such as albumin and alpha-fetoprotein, and inflammatory cytokines, such as IFN-g and TNF-α. In this review, we provide an overview of the recent advancements in biosensors for tissue engineering applications.

Matched MeSH terms: Antibodies/chemistry*

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