. 2019 Mar 8;28(1):99–100. doi: 10.1159/000496961
Med Princ Pract. 2019 Mar 8;28(1):99.
Med Princ Pract. 2019 Mar 8;28(1):99.
Periodontal diseases are a group of chronic infections that destroy tissues surrounding and supporting the teeth. There are no data on the anaerobes associated with periodontal infections in Kuwait. The objective of this study was to investigate the target anaerobes associated with periodontal diseases in Kuwait. Patients with chronic periodontitis, seen at the Kuwait University Dental Clinics, were recruited into this study during a period of 15 months. Samples were collected using Gutta Percha Points directly from inside the gingival pockets and transported in sterile Ringer's solution and prereduced thioglycolate broth to the laboratory. One set of samples was cultured on appropriate media, and the other set was used for polymerase chain reaction (PCR) assays. DNA sequencing and semiquantitative PCR were performed for Prevotella spp. only. A total of 30 patients stratified according to the severity of the disease; mild (n = 3), moderate (n = 14), and severe (n = 13), were studied. Thirty-one healthy individuals were included as controls. Thirty percent of the patients were between 50 and 59 years old. The female-to-male ratio was 1: 2. The ratio of Kuwaitis to non-Kuwaitis was 1: 3.3. Aggregatibacter actinomycetemcomitans were found in 23.3 versus 16.1% of the cases and controls, respectively (p > 0.05). All 30 (100%) patients harbored Fusobacterium spp. compared to 93.5% of the controls (p > 0.05). Porphyromonas gingivalis was detected in 33.3% of the patients versus 6.4% of the controls (p < 0.0001). Tannerella forsythia was detected in 83.3% of the patients versus 51.6% of the controls (p < 0.0001). Parvimonas micra was detected in 90% of the patients versus 51.6% of the controls (p < 0.0001), and Treponema denticola was detected in 70% of the patients versus 29% of the controls (p < 0.0001). Prevotella spp. were detected in all patients and controls; the majority was P. nigrescens detected in 70 and 100% of the patients and controls, respectively. There was no significant difference in DNA concentrations of Prevotella spp. between patients and controls by semiquantitative PCR assay. In conclusion, all target anaerobes, except Aggregatibacter spp., Prevotella spp., and Fusobacterium spp., were significantly associated with periodontitis in our study. P. gingivalis was the most strongly associated anaerobe with periodontal disease, while Prevotella spp. appear to be normal passengers.
Dr. Wafaa Jama (Supervisor)
Prof. Vincent Rotimi (Co-Supervisor)
Med Princ Pract. 2019 Mar 8;28(1):100.
Med Princ Pract. 2019 Mar 8;28(1):100.
Human coronavirus OC43 (HCoV-OC43) is a respiratory virus that usually causes common cold. However, it has the potential to cause severe infection in young children and immunocompromised adults. Both SARS-CoV and MERS-CoV were shown to express proteins with the potential to evade early innate immune responses. However, the ability of HCoV-OC43 to antagonize the intracellular antiviral defenses has not yet been investigated. The potential role of the HCoV-OC43 structural (M and N) and accessory proteins (ns2a and ns5a) in the alteration of antiviral gene expression was investigated in this study. HCoV-OC43 M, N, ns2a, and ns5a proteins were expressed in human embryonic kidney 293 (HEK-293) cells before challenging with Sendai virus (SeV). The Human Antiviral Response PCR array was used to profile the antiviral gene expression in HEK-293 cells, whereas the effect of HCoV-OC43 proteins on the transcriptional activation of antiviral response elements was assessed by measuring the levels of firefly luciferase expressed by a reporter vector under the control of interferon-stimulated response element (ISRE), interferon-beta (IFN-β) promoter, or nuclear factor kappa B response element (NF-κB-RE). Influenza A NS1 protein was used as a positive control for IFN antagonism. Over 30 genes were downregulated in the presence of one of the HCoV-OC43 proteins; e.g., mitogen-activated protein kinases, Toll-like receptors, interferons, interleukins, and signalling transduction proteins. The transcriptional activity of ISRE, IFN-β promoter, and NF-κB-RE was significantly reduced in the presence of HCoV-OC43 ns2a, ns5a, M, or N protein, as there was a sharp fall in the firefly luciferase levels following the challenge of cells with SeV, IFN-α, or TNF-α. Our findings suggest that similarly to SARS-CoV and MERS-CoV, HCoV-OC43 can downregulate the transcription of genes critical for the activation of different antiviral signalling pathways. Further studies are needed to confirm the role of HCoV-OC43 structural and accessory proteins in antagonizing antiviral gene expression.
Dr. Wassim Chehadah (Supervisor)
Med Princ Pract. 2019 Mar 8;28(1):100.
Candida species are a major cause of healthcare-associated infections. Rapid identification and characterization of Candida species are needed for effective preventive and management strategies. This study had three main objectives: (i) to develop molecular methods for rapid detection of several Candida spp. (C. albicans and C. dubliniensis, C. parapsilosis-complex and C. glabrata-complex) by multiplex polymerase chain reaction (PCR) assays; (ii) to determine genotypic heterogeneity/relatedness among clinical isolates of C. albicans-complex, C. parapsilosis-complex, and C. glabrata-complex to study the origin of nosocomial Candida infections and to develop new strategies for treatment, control, and prevention of candidemia cases in Kuwait; and (iii) to perform antifungal susceptibility testing and to determine clade specificity and molecular basis of drug resistance among Candida spp. isolates. Major findings included: (i) two duplex PCR and two multiplex PCR assays were developed for rapid identification and differentiation of individual species among several Candida species complexes; (ii) fingerprinting of C. albicans isolates from candidemia patients by MLST and AFLP analyses coupled with epidemiological data revealed that individual patients were infected with unique strains, thus ruling out hospital transmission of the infection; (iii) fingerprinting of C. dubliniensis isolates revealed five ITS haplotypes and only 16 MLST-based DST, whereby all isolates resistant to 5-FC belonged to ITSH4/DST14 and contained S29L mutation in CdFCA1; (iv) an increasing trend in the resistance to fluconazole among C. parapsilosis isolates and the detection of endemic genotypes in neonatal intensive care units (ICUs) during fingerprinting by MLMT; (v) fingerprinting of clinical C. orthopsilosis isolates identified 3 ITS haplotypes and 7 amplified fragment-length polymorphism genotypes including one genotype that has persisted over several years; and (vi) fingerprinting of C. glabrata strains showed that fingerprinting by only two loci had the same discriminatory power as the combined data from 8 loci. In conclusion, several duplex/multiplex PCR assays have been developed for the detection of different members of various Candida species complexes. Our data showed that related strains of C. albicans exist and fingerprinting by MLST alone may complicate hospital infection control measures during outbreak investigations. All 5-FC-resistant C. dubliniensis isolates belonged to a single genotype. Fingerprinting of C. parapsilosis isolates showed presence of endemic genotypes in some neonatal ICUs and suggested their transmission to other patients hospitalized in the same unit. We also developed a highly discriminatory fingerprinting method based on only 2 loci for strain differentiation of C. glabrata isolates. The study contributes to our understanding of the molecular epidemiology of Candida infections in Kuwait.
Prof. Zia Uddin Khan (Supervisor)
Prof. Suhail Ahmad (Co-Supervisor)