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Fulltext Conventional rapid latex agglutination in estimation of von Willebrand factor: method revisited and potential clinical applications

Mahat M, Abdullah WZ, Hussin CM

J Immunol Res, 2014;2014:850810.
PMID: 25759835 DOI: 10.1155/2014/850810

Measurement of von Willebrand factor antigen (VWF : Ag) levels is usually performed in a specialised laboratory which limits its application in routine clinical practice. So far, no commercial rapid test kit is available for VWF : Ag estimation. This paper discusses the technical aspect of latex agglutination method which was established to suit the purpose of estimating von Willebrand factor (VWF) levels in the plasma sample. The latex agglutination test can be performed qualitatively and semiquantitatively. Reproducibility, stability, linearity, limit of detection, interference, and method comparison studies were conducted to evaluate the performance of this test. Semiquantitative latex agglutination test was strongly correlated with the reference immunoturbidimetric assay (Spearman's rho = 0.946, P < 0.001, n = 132). A substantial agreement (κ = 0.77) was found between qualitative latex agglutination test and the reference assay. Using the scoring system for the rapid latex test, no agglutination is with 0% VWF : Ag (control negative), 1+ reaction is equivalent to <20% VWF : Ag, and 4+ reaction indicates >150% VWF : Ag (when comparing with immunoturbidimetric assay). The findings from evaluation studies suggest that latex agglutination method is suitable to be used as a rapid test kit for the estimation of VWF : Ag levels in various clinical conditions associated with high levels and low levels of VWF : Ag.
Fulltext The antinociceptive effects of tualang honey in male sprague-dawley rats: a preliminary study

Aziz CB, Ismail CA, Hussin CM, Mohamed M

J Tradit Complement Med, 2014 Oct;4(4):298-302.
PMID: 25379476 DOI: 10.4103/2225-4110.139115

Tualang honey ( Fēng Mì) is known to have anti-inflammatory property, but its antinociceptive property has not been extensively investigated. In this study, we examined the preemptive effects on administering different doses of Tualang honey and prednisolone on the nociceptive response in male Sprague-Dawley rats. Thirty-five male Sprague-Dawley rats were randomized into five groups (n = 7) and each group received either distilled water, Tualang honey (0.2, 1.2 or 2.4 g/kg) or prednisolone (10 mg/kg) for 10 days. The response to noxious thermal stimulus was assessed using tail flick test on Day 10. The well-being of the rats was also assessed by monitoring their food intake and body weight. Data were analyzed using one-way Analysis of Variance (ANOVA) with post-hoc Scheffe's test and P value less than 0.05 was considered significant. In tail flick test, the tail flick latency time was significantly higher in the groups that received 1.2 g/kg and 2.4 g/kg of Tualang honey and 10 mg/kg of prednisolone, compared to the control group (P < 0.05). There was significant reduction in the total food pellet intake in the groups receiving prednisolone and Tualang honey (1.2 g/kg and 2.4 g/kg) compared to controls; however, the body weight gain was only significantly reduced in the prednisolone group. All the parameters were not significantly affected in the group receiving 0.2 g/kg of Tualang honey. In conclusion, preemptive administration of Tualang honey (1.2 g/kg and 2.4 g/kg) and prednisolone (10 mg/kg) had reduced the pain responses. The reduced weight gain in the prednisolone group is an unwanted effect due to its metabolic and central actions. Further studies are required to confirm the antinociceptive effects and elucidate the mechanism of antinociceptive action of Tualang honey in the rats.
Clinical features of allergic rhinitis and skin prick test analysis based on the ARIA classification: a preliminary study in Malaysia

Asha'ari ZA, Yusof S, Ismail R, Che Hussin CM

Ann Acad Med Singap, 2010 Aug;39(8):619-24.
PMID: 20838703

INTRODUCTION: Allergic rhinitis (AR) is a prevalent disease worldwide but is still underdiagnosed in many parts of Asia. We studied the clinical profiles of AR patients in our community based on the new ARIA classification and investigated the aetiological allergens using a skin prick test.
MATERIALS AND METHODS: In 2008, 142 newly diagnosed patients with AR were seen and underwent skin prick testing with 90 patients completing the study.
RESULTS: Intermittent mild and moderate/severe AR were evident in 10% and 21.1% of the patients, while persistent mild and moderate/severe were seen in 20% and 48.9%, respectively. Rhinitis and asthma co-morbidity occurred in 28.8% with asthma incidence significantly higher in persistent AR (P = 0.002). There was no significant association between AR severity, city living and asthma co-morbidity. Nasal itchiness and sneezing were the main presenting complaints and were more common in intermittent AR (P <0.05). Sleep disturbance was associated with moderate-severe AR (P <0.05). Polypoidal mucosa was associated with asthma co-morbidity (P <0.05). Monosensitivity reaction occurred in 12.2% of patients and was associated with fungi sensitivity (P <0.05). Majority of patients were oligosensitive (52.8%) and polysensitive (34.4%) and were significantly associated with moderate-severe persistent AR (P <0.01). The highest positive skin prick reaction and the largest average wheal diameter were for the house dust mites and cat allergen (P <0.05).
CONCLUSION: Our results reflected the AR profiles in our country, which was comparable with typical profiles of the neighbouring country and other Mediterranean countries with a similar temperate climate.
Anti-CCP antibodies: A better diagnostic tool for rheumatoid arthritis

Dollah R, Yaacob A, Ismail AH, Che Hussin CM

International Medical Journal (Tokyo), 2011;18(1):53-57.

Objective: The assay for anti-cyclic citrullinated peptide (anti-CCP) antibody is a recent test of interest due to low diagnostic specificity of rheumatoid factor in the diagnosis of rheumatoid arthritis. To determine the sensitivity and specificity of anti-CCP antibodies in rheumatoid arthritis patients using American College of Rheumatology (ACR) criteria as a gold standard.
Design: A cross sectional study was conducted from January 2008 to December 2008 on 100 patients with rheumatoid arthritis and 153 patients with arthritis or arthralgia but not fulfilling ACR criteria for rheumatoid arthritis.
Materials and Methods: Serum from each subject was tested for anti-CCP antibodies and IgM rheumatoid factor (IgM RF) by an enzyme linked immunosorbent assay (ELISA). Sensitivity and specificity of the tests were evaluated using the ACR criteria as the gold standard. Data was analyzed using SPSS version 12.0 software. The association between the two categorical variables were tested using chi-square test. The differences between the two groups were tested using independent t-test. Results: The sensitivity of anti-CCP antibodies was 71% with 94.8% of specificity. For rheumatoid factor the sensitivity was 85% and the specificity was 74.5%. Positive predictive value for anti-CCP antibodies was 89.9% whereas for rheumatoid factor it was 68.5%. The sensitivity of ACR criteria with inclusion of anti- CCP antibodies was 100% and specificity of 94.8% (95%CI: 91.3%, 98.2%).
Conclusion: Anti-CCP antibody has a higher diagnostic accuracy compared to the RF. Anti-CCP antibody is a useful laboratory marker to support the diagnosis of rheumatoid arthritis and to differentiate patients with rheumatoid arthritis from rheumatoid arthritis-like symptoms. Therefore, it is suggested that anti-CCP antibodies should be included as another laboratory supportive criterium besides RF test in ACR criteria in the diagnosis of RA.
Study site: family medicine clinic, rheumatology clinic and immunology laboratory of Hospital Universiti Sains Malaysia (HUSM), Kubang Kerian, Kelantan, Malaysia
Fulltext CD3+CD4+gp130+ T Cells Are Associated With Worse Disease Activity in Systemic Lupus Erythematosus Patients

Mohd Shukri ND, Farah Izati A, Wan Ghazali WS, Che Hussin CM, Wong KK

Front Immunol, 2021;12:675250.
PMID: 34149710 DOI: 10.3389/fimmu.2021.675250

The receptors for IL-35, IL-12Rβ2 and gp130, have been implicated in the inflammatory pathophysiology of autoimmune diseases. In this study, we set out to investigate the serum IL-35 levels and the surface levels of IL-12Rβ2 and gp130 in CD3+CD4+, CD3+CD4─ and CD3─CD4─ lymphocyte subpopulations in systemic lupus erythematosus (SLE) patients (n=50) versus healthy controls (n=50). The potential T cell subsets associated with gp130 transcript (i.e. IL6ST) expression in CD4+ T cells of SLE patients was also examined in publicly-available gene expression profiling (GEP) datasets. Here, we report that serum IL-35 levels were significantly higher in SLE patients than healthy controls (p=0.038) but it was not associated with SLEDAI-2K scores. The proportions of IL-12Rβ2+ and gp130+ cells in SLE patients did not differ significantly with those of healthy controls in all lymphocyte subpopulations investigated. Essentially, higher SLEDAI-2K scores were positively correlated with increased proportion of gp130+ cells, but not IL-12Rβ2+ cells, on CD3+CD4+ T cells (r=0.425, p=0.002, q=0.016). Gene Set Enrichment Analysis (GSEA) of a GEP dataset of CD4+ T cells isolated from SLE patients (n=8; GSE4588) showed that IL6ST expression was positively associated with genes upregulated in CD4+ T cells vs myeloid or B cells (q<0.001). In an independent GEP dataset of CD4+ T cells isolated from SLE patients (n=9; GSE1057), IL6ST expression was induced upon anti-CD3 stimulation, and that Treg, TCM and CCR7+ T cells gene sets were significantly enriched (q<0.05) by genes highly correlated with IL6ST expression (n=92 genes; r>0.75 with IL6ST expression) upon anti-CD3 stimulation in these SLE patients. In conclusion, gp130 signaling in CD3+CD4+ T cell subsets may contribute to increased disease activity in SLE patients, and it represents a promising therapeutic target for inhibition in the disease.
Fulltext Increased IL-23R+ Th Cells Population Exhibits Higher SLEDAI-2K Scores in Systemic Lupus Erythematosus Patients

Izati AF, Mohd Shukri ND, Wan Ghazali WS, Che Hussin CM, Wong KK

Front Immunol, 2021;12:690908.
PMID: 34484186 DOI: 10.3389/fimmu.2021.690908

The IL-23/IL-17 axis plays causative roles in the development and progression of systemic lupus erythematosus (SLE). However, it remains unclear if the IL-17RA+ and IL-23R+ T helper (Th) cells populations are associated with the serum IL-17 and IL-23 levels, or with the immunological parameters and disease activities in SLE patients. Herein, we examined the proportion of IL-17RA+ and IL-23R+ Th cells and serum levels of IL-17 and IL-23 in established SLE patients (n = 50) compared with healthy controls (n = 50). The associations of these interleukins and their receptors with immunological parameters [anti-nuclear antibody (ANA), anti-dsDNA antibody, and C-reactive protein (CRP)] and SLE disease activity (SLEDAI-2K scores) in SLE patients were assessed. CD3+CD4+ Th cells of SLE patients demonstrated significantly elevated IL-17RA+ (p = 1.12 x 10-4) or IL-23R+ (p = 1.98 x 10-29) populations compared with the healthy controls. Serum IL-17 levels were significantly lower in SLE patients compared with the healthy controls (p = 8.32 x 10-5), while no significant difference was observed for the IL-23 serum levels between both groups. IL-23R+ Th cells population was significantly associated with higher SLEDAI-2K scores (p = 0.017). In multivariate analysis, the proportion of IL-23R+ Th cells remained significantly associated with higher SLEDAI-2K scores independent of prednisolone intake (p = 0.027). No associations were observed between the interleukin parameters (i.e., IL-17, IL-23, IL-17RA+ Th cells, and IL-23R+ Th cells) with ANA, anti-dsDNA, and CRP status, suggesting that the IL-17/IL-23 axis acts independently of these immunological parameters. In conclusion, our results support that therapeutic inhibition of the IL-23/IL-17 axis receptors on Th cells, particularly IL-23R, is potentially relevant in SLE patients.
Major depressive disorder patients on antidepressant treatments display higher number of regulatory T cells

Mohd Ashari NS, Mohamed Sanusi SNF, Mohd Yasin MA, Che Hussin CM, Wong KK, Shafei MN

Malays J Pathol, 2019 Aug;41(2):169-176.
PMID: 31427552

INTRODUCTION: Regulatory T cell (Treg) is a subtype of T lymphocyte that plays a crucial role in establishing immunologic self-tolerance and maintaining immune homeostasis. In this study, we set out to investigate the percentage and absolute count of Tregs in major depressive disorder (MDD) patients and their correlation with disease severity.
MATERIALS & METHODS: This is a case-control study consisting of 47 MDD patients and 47 healthy controls. MDD patients were treated with antidepressant drugs according to their physician's choice. The severity of MDD was assessed using Beck Depression Inventory (BDI) and Montgomery-Asberg Depression Rating Scale (MADRS) at the time of recruitment. Healthy controls completed the Depression Anxiety Scoring System (DASS21) questionnaire to ensure they were in good mental health without history of MDD. The percentage and absolute count of CD4+ CD25+ Tregs and CD4+ CD25+ FOXP3+ Tregs were identified by multiparameter flow cytometry.
RESULTS: The percentage and absolute count of CD4+ CD25+ Treg cells were significantly higher in MDD patients than in healthy controls (P<0.001, in both cases). Likewise, the percentage and absolute count of CD4+ CD25+ FOXP3+ Treg cells were also significantly higher in MDD patients compared to healthy controls (P=0.003 and P=0.002, respectively). However, there was no significant correlation between the percentage and absolute count of CD4+ CD25+ Treg and CD4+ CD25+ FOXP3+ Treg cells with BDI or MADRS score.
CONCLUSIONS: Our results suggest that antidepressant treatments contributed to an upregulation of Tregs in MDD patients.