Analysis of IFN-kappa expression in pathologic skin conditions: downregulation in psoriasis and atopic dermatitis.

Interferon-kappa (IFN-kappa) is a type I IFN expressed by keratinocytes, monocytes and dendritic cells (DCs). In human keratinocytes, it is produced in response to double-stranded RNA (dsRNA) and other IFNs and protects from viral infections. In monocytes and DCs, IFN-kappa induces tumor necrosis factor-alpha (TNF-alpha) and interleukin-10 (IL-10) and inhibits lipopolysaccharide (LPS)-induced IL-12. In this study, we evaluated IFN-kappa expression in skin lesions of patients with common immune-mediated inflammatory disorders using immunohistochemical techniques. IFN-kappa was not detectable in healthy skin but was strongly expressed in allergic contact dermatitis and lichen planus-affected skin. IFN-kappa was localized mainly in basal and suprabasal keratinocytes and in some leukocytes infiltrating the dermis. In contrast, IFN-kappa expression in psoriatic or atopic dermatitis (AD) pidermis was weak and detectable in only 2 of 5 patients examined. Consistently, cultured keratinocytes and monocytes obtained from psoriatic and AD patients expressed null or low levels of IFN-kappa in response to IFN-gamma, which strongly upregulates IFN-kappa in normal keratinocytes. IFN-kappa accumulated in keratinocyte cytoplasm and plasma membrane, and only limited amounts were released extracellularly. Soluble IFN-kappa did not influence keratinocyte proliferation or chemokine and membrane molecule expression, and only its membrane-associated form activated IFN-stimulated response element (ISRE) signaling. Given the difference in IFN-kappa expression levels in the skin disorders examined, IFN-kappa presence or deficiency might have different pathogenetic consequences depending also on other disease-specific intrinsic alterations.     

Nitric oxide donors suppress chemokine production by keratinocytes in vitro and in vivo

Nitric oxide (NO) is involved in the modulation of inflammatory responses. In psoriatic skin, NO is highly produced by epidermal keratinocytes in response to interferon-gamma and tumor necrosis factor-alpha. In this study, we investigated whether the NO donors, S-nitrosoglutathione (GS-NO) and NOR-1, could regulate chemokine production by human keratinocytes activated with interferon-gamma and tumor necrosis factor-alpha. In addition, we studied the effects of the topical application of a GS-NO ointment on chemokine expression in lesional psoriatic skin. NO donors diminished in a dose-dependent manner and at both mRNA and protein levels the IP-10, RANTES, and MCP-1 expression in keratinocytes cultured from healthy patients and psoriatic patients. In contrast, constitutive and induced interleukin-8 production was unchanged. GS-NO-treated psoriatic skin showed reduction of IP-10, RANTES, and MCP-1, but not interleukin-8 expression by keratinocytes. Moreover, the number of CD14(+) and CD3(+) cells infiltrating the epidermis and papillary dermis diminished significantly. NO donors also down-regulated ICAM-1 protein expression without affecting mRNA accumulation in vitro, and suppressed keratinocyte ICAM-1 in vivo. Finally, NO donors inhibited nuclear factor-kappa B and STAT-1, but not AP-1 activities in transiently transfected keratinocytes. These results define NO donors as negative regulators of chemokine production by keratinocytes.     

Distinct delayed T-cell response to beta-methasone and penicillin-G in the same patient

BACKGROUND: Multiple drug allergy syndrome is a clinical condition characterized by reactions against more than one different class of, both pharmacologically and structurally, unrelated drugs. Scanty data are available to date about a multiple drug delayed hypersensitivity syndrome. Our aim was to report the case of a delayed reaction to both beta-methasone (beta-MT) and penicillin-G (pen-G) occurring in the same patient, and analyse beta-MT- and pen-G-specific T-cell Lines (TCLs) with regard to their specificity, phenotype and cytokine profile. METHODS: We generated two drug-specific TCLs from biopsies at the site of positive intradermal reactions, and analysed their immunophenotype, T-cell receptor Vbeta (TCR-Vbeta) domains expression and cytokine profile. RESULTS: We demonstrated the specificity of the T cells isolated from positive intradermal test reactions to pen-G and beta-MT through the strict dose-dependent proliferation in response to drug-pulsed autologous antigen presenting cells. Fluorescence activated cell sorter (FACS) analysis revealed a predominance of CD4+ cells in the inflammatory cell infiltrate of intradermal test with beta-MT, while a predominance of CD8+ T cells in the site of delayed reaction to pen-G was found. The drug specific CD4+ and CD8+ T cells were heterogeneous, with regard to TCR-Vbeta usage. CD8+ pen-G-TCL displayed a preferential T helper 2 (Th2) profile, while a substantially heterogeneous pattern of cytokine production characterized specific beta-MT TCL. CONCLUSION: The study describes the coexistence in the same patient of a delayed hypersensitivity to both penicillin G and beta-MT, driven, respectively, by pen-G-specificTh2-skewed CD8+ and beta-MT specificTh0 CD4+ T cells. This case further support the existence of a multiple drug allergy syndrome also for delayed hypersensitivity.     

Pneumococcal infection in hospitalized patients: a four-year study in Malaysia

A total of 90 cases of pneumococcal infections were identified at a major referral hospital in Kuala Lumpur, Malaysia during a study period of four years. Pneumonia was the most common clinical presentation (41 cases) followed by meningitis (19 cases). Of 48 patients who were followed-up during the microbiology consultation round, 11 died, 9 were children below two years old. Capsular typing was carried out on 57 strains of Streptococcus pneumoniae isolated from blood and body fluids of 43 children and 14 adults. 38 strains isolated from pharyngeal specimens were also typed. Types 6A (11 strains), 6B (7 strains), 14 (8 strains) and 19A (8 strains) predominated in children. The strains from older patients comprised 3 isolates from cerebrospinal fluid (types 18B, 6B and 14), five from blood (4 strains, type 1 and 1 strain, type 4) and six from pus (1 strain, type 14, 3 strains type 23F and 2 strains type 34). The isolates from pharyngeal specimens belonged to capsular type similar to those implicated in infections. 90% of the types reported in this study are included in the 23 valent pneumococcal vaccines. Minimum inhibitory concentrations of penicillin, cefuroxime, chloramphenicol and rifampicin were determined for selected strains. 4.1% of isolates were resistant to penicillin (3/74), 4.5% to cefuroxime (2/44), 6.5% to chloramphenicol (3/46) and 14.6% to rifampicin (6/41).     

Surgical treatment of bisphosphonate-associated osteonecrosis of the jaw: Technical report and follow up of 21 patients

IntroductionBisphosphonates are used to reduce skeletal related events in patients with bone consuming diseases such as osteoporosis and bone metastases. However recently there has been an increased awareness of bisphosphonate-associated necrosis of the jaws (BP-ONJ). Many authors propose conservative management in these cases but invariably the problem is not treated successfully allowing the bone defect to worsen. Recently there has been a move to treat this problem surgically. The aim of this retrospective study was to provide a surgical solution for patients suffering from BP-ONJ.Materials and methodsAll patients presenting with BP-ONJ were treated with bone debridement of the affected area and multilayer wound closure. The considered variables were: gender, age, underlying diagnosis, type of bisphosphonate (BP) used, duration of bisphosphonate use, route of administration, location of the osteonecrosis, clinical symptoms, association with dental treatment and surgical outcome.ResultsNineteen cases of a total of 21 demonstrated no recurrence of osteonecrosis during follow up (Mean 16 months – Range 12–24 months). One patient with a bilateral defect showed a dehiscence on one side and a small fistula on the contralateral side 6 weeks post-operatively and required revision surgery. Another patient developed a fistula after 4 weeks that was treated successfully with antibiotics and curettage. No patients had evidence of exposed bone, bland mucosa nor pain at the surgical site.ConclusionThe technique described can be recommended for patients with BP-ONJ if a conservative treatment fails.     

Azathioprine-related bone marrow toxicity and low activities of purine enzymes in patients with rheumatoid arthritis

Objective. Azathioprine (AZA) metabolism largely parallels the endogenous purine pathways. To date, thiopurine methyltransferase (TPMT) deficiency has been reported as a cause of AZA-related bone marrow toxicity in 1 patient with rheumatoid arthritis (RA). We therefore studied purine enzyme activities in 3 patients with RA who experienced AZA-related bone marrow toxicity. Methods. Lymphocyte activity of purine nucleoside phosphorylase and 5′-nucleotidase (5NT) and erythrocyte activity of TPMT, key enzymes in thiopurine catabolism, were measured in 3 RA patients who had experienced AZA-related bone marrow toxicity and in 16 RA patients without signs of toxicity despite at least 6 months of treatment with AZA. Results. Two patients with AZA-related bone marrow toxicity were found to have a TPMT deficiency, 1 partial and 1 total. In the third patient, 5NT activity was found to be well below the lowest level observed in the control subjects. Conclusion. All 3 patients with severe AZA-related bone marrow toxicity had abnormal purine enzyme activities. Deficiency of purine enzymes, including TPMT and 5NT, may be a cause of AZA-related bone marrow toxicity in patients with RA.     

The capacity of the third-stage larvae of Oesophagostomum bifurcum to survive adverse conditions

Human infections with the intestinal nematode Oesophagostomum bifurcum are commonly found in the Sudan savannah of northern Togo and Ghana. Apparently, the long and hot dry season in this region does not prevent transmission, which is believed to take place through ingestion of the infective, third-stage larvae (L3). Oesophagostomum L3 cultured from human stools, unlike the larvae of Necator americanus, were shown to survive desiccation. In addition, 93% of the O. bifurcum L3 frozen for 24 h at -15 degrees C regained motility when brought back into ambient temperatures. The L3 also survived the acidity of an artificial mixture made to resemble the gastric juices of humans. Desiccated larvae could even be rehydrated in this mixture, indicating the possibility of dust-borne infections. The sturdiness of the L3 is likely to contribute to the high transmission intensity in northern Togo and Ghana.     

Anatomy of the arcus tendineus fasciae pelvis in females

Because of its proximity to the urethra, the anterior part of the arcus tendineus fasciae pelvis (ATFP) may be used in urethrosuspension procedures for urinary stress incontinence. In this study, 10 embalmed female cadaver hemipelves were dissected and their gross anatomy described. In females, the ATFP is a condensation of the endopelvic fascia. The anterior attachment of the ATFP is to the caudal inner surface of the body of the pubic bone at a site averaging 4 mm lateral to the pubic symphysis and covering an average area of 53 mm(2). Posteriorly, it attaches to the medial surface of the ischial spine. In nine of the 10 hemipelves the first anterior centimeters of the ATFP have a clear lateral fixation either to the lateral part of the levator ani muscle (n = 1), to the fascia covering the obturator internus muscle (n = 7), or to the obturator membrane (n = 1). Medially from the ATFP derives a 2-3-cm long flat fibrous attachment to the posterolateral aspect of the urethra. In eight unembalmed cadavers, the ATFP gave way at a pulling force of 8.2 kg (range = 3.5-11.5 kg). The ATFP resists caudal movement of the proximal anterior vaginal wall and the urethra in the upright posture and, therefore, may be suitable for urethrosuspension procedures.