The human microbiome in immunobullous disorders and lichen planus.

For several decades, there has been a significant growth in the incidence of autoimmune diseases. Studies indicate that genetic factors may not be the only trigger for disease development and that dysbiosis of the microbiome may be another mechanism involved in the pathogenesis of autoimmune diseases. The role of the microbiome in the development of common skin disorders such as psoriasis, atopic dermatitis, acne and rosacea is increasingly well understood. However, few studies have focused on lichen planus and the rare acquired immunobullous diseases, both mucocutaneous groups of disorders linked to skin, oral and gut microbiomes. This review provides an insight into the current understanding of how the microbiome may contribute to the development of autoimmunity and to the maintenance and exacerbation of acquired immunobullous and lichenoid diseases. These mechanisms may have implications for future preventive and therapeutic approaches.

The perceived impact of Covid-19 on periodontal practice in the United Kingdom: A questionnaire study.

INTRODUCTION: COVID-19 has impacted dentistry in unprecedented ways. OBJECTIVE: The following research aimed to investigate the impact of the COVID-19 pandemic on periodontal practice in the United Kingdom using the COM-B (Capability Opportunity Motivation-Behaviour) model as the basis for a questionnaire. BASIC RESEARCH DESIGN: An online survey link was sent to all members of the British Society of Periodontology and Implant Dentistry. A total of 358 responses were received and analysed. RESULTS: The great majority of participants thought that the pandemic had an impact on their profession, while only 4.7 % had no concerns. The main worries related to financial concerns and ability to provide appropriate levels of care. More than 80 % of respondents agreed that their establishment was compliant with infection control procedures. Some participants felt benefits mainly in terms of more time for CPD activities. It was felt that some of the changes needed will need to be sustained long-term. CONCLUSIONS: Respondents were generally worried. However, they perceived they had the physical and psychological ability to effect changes to their practice, higher than the physical and social opportunities that they were afforded. Although the COVID-19 pandemic is causing profound changes and worries for the profession of Periodontology, clinicians are clear about their capability to control the situation and feel they have the motivation to make the required changes. CLINICAL SIGNIFICANCE: COVID-19 has presented clinicians with novel challenges. Investigating the professional response to change and expected impact is of interest in the current climate as we navigate the 'new normal'. Assessing the results could be useful in informing support strategies moving forward.

Centronuclear Myopathy with Abundant Nemaline Rods in a Japanese Black and Hereford Crossbred Calf.

Histopathological examination was performed on skeletal and diaphragmatic muscles from an 8-month-old male crossbred calf showing abnormal gait and tremor of the hindlimbs. There were numerous round fibres with centrally placed nuclei forming nuclear chains in longitudinal sections, associated with interstitial fibrosis or adipose tissue infiltration. On nicotinamide adenine dinucleotide tetrazolium reductase (NADH-TR) staining, some muscle fibres in severe lesions showed a spoke-like appearance due to a radial arrangement of sarcoplasmic strands. Additionally, increased NADH-TR activity in the subsarcolemmal structures, appearingas ring-like or necklace-like forms, were observed. Transmission electron microscopy revealed dilated sarcoplasmic reticulum and variably shaped electron-dense inclusions consisting of myofibrillar streams. Another prominent feature was the existence of numerous nemaline rods within muscle fibres; these were stained red by Gomori's trichrome stain. Immunohistochemistry revealed that the nemaline rods showed strong immunoreactivity with α-actinin and desmin antibodies. Electron microscopically, these structures were composed of dense-homogeneous material and continuous with the Z disk. The case was diagnosed as centronuclear myopathy with increased nemaline rods.

Retrospective and prospective studies of hepatitis B virus reactivation in malignant lymphoma with occult HBV carrier.

BACKGROUND: In surface antigen of hepatitis B virus (HBsAg)-positive carrier for anticancer treatment of malignant lymphoma, it is well recognized that reactivation of hepatitis B virus (HBV) occasionally occurs. However, there have been only a few studies of HBV reactivation in serum HBsAg-negative and hepatitis B core antigen (HBcAb)-positive occult HBV carriers. We looked at both retrospective and prospective studies to determine the prevalence, clinical course and risk factor of HBV reactivation during chemotherapy in lymphoma patients. PATIENTS AND METHODS: Forty-eight of 127 (37.8%) lymphoma patients were HBsAg negative and HBcAb positive, and 24 of these patients were then given liver function tests and HBsAg tests monthly and serum HBV DNA every 3 months. RESULTS: HBV reactivation was observed in two patients (4.1%) who had received intensive chemotherapy including steroid and rituximab. Immediate administration of entecavir therapy after elevation of HBV DNA level was conducted, and this resulted in reduction of it and improvement of liver function test. CONCLUSIONS: Rituximab plus steroid-containing regimens may increase the risk of HBV reactivation in HBsAg-negative and HBcAb-positive lymphoma patients. More ambitious prospective studies are required to establish clinically useful or cost-effective follow-up methods for control of HBV reactivation in lymphoma patients with occult HBV infection.

Psychological interventions to improve adherence to oral hygiene instructions in adults with periodontal diseases.

BACKGROUND: Adherence to oral hygiene is an important aspect of the treatment of periodontal disease. Traditional educational interventions have been shown to be of little value in achieving long term behaviour change. OBJECTIVES: The aim of this review was to determine the impact of interventions aimed to increase adherence to oral hygiene instructions in adult periodontal patients based on psychological models and theoretical frameworks. This review considered the following outcomes: Observational measures of oral health related behaviour Self reported oral health related behaviours, beliefs and attitudes towards oral health related behaviour Clinical markers of periodontal disease. SEARCH STRATEGY: The Cochrane Oral Health Group's Trials Register (2005), the Cochrane Central Register of Controlled Trials (CENTRAL) (The Cochrane Library 2004, Issue 4), MEDLINE (from 1966 to December 2004), EMBASE (from 1980 to December 2004), PSYCHINFO (from 1966 to December 2004), Ingenta (from 1998 to December 2004) and CINAHL (from 1966 to December 2004). Reference lists from relevant articles were searched and the authors of eligible trials were contacted to identify trials and obtain additional information. No language restriction was applied. SELECTION CRITERIA: Randomised controlled trials testing the effectiveness of interventions based on psychological models compared with educational, attention or no active intervention controls to improve adherence to oral hygiene in adults with either gingivitis or periodontitis. DATA COLLECTION AND ANALYSIS: Titles and abstracts of studies that were potentially relevant to the review were independently screened by two review authors. Those that were clearly ineligible were rejected. For the remaining studies, the full paper was reviewed by two review authors and where necessary further information was sought from the author to verify eligibility. Included studies were assessed on their quality using standard criteria. MAIN RESULTS: The review identified four studies (including 344 participants) in which a psychological model or theory had been explicitly used as the basis for the design of the intervention. The overall quality of trials was low. Due to the hetereogeneity between studies, both in terms of outcome measures and psychological models adopted, a meta-analysis was not possible. The four studies adopted four different theoretical frameworks, though there was some overlap in that three of the studies incorporated elements of Operant and Classical Conditioning. Psychological interventions resulted in improved plaque scores in comparison to no intervention groups, and in one study in comparison to an attention control group. One study found decreased gingival bleeding in the active intervention group but no change in pocket depth or attachment loss after 4 months. Psychological interventions were associated with improved self reported brushing and flossing in both studies which assessed these behaviours. Only one study explored the impact of psychological interventions on beliefs and attitudes, the psychological intervention, in comparison to educational and no intervention controls, showed improved self efficacy beliefs in relation to flossing, but no effect on dental knowledge or self efficacy beliefs in relation to tooth brushing. AUTHORS' CONCLUSIONS: There is tentative evidence from low quality studies that psychological approaches to behaviour management can improve oral hygiene related behaviours. However, the overall quality of the included trials was low. Furthermore, the design of the interventions was weak and limited, ignoring key aspects of the theories. Thus, there is a need for greater methodological rigour in the design of trials in this area.

Musculoskeletal neck and back pain in undergraduate dental students at a UK dental school - a cross-sectional study.

Objective Limited data exist on musculoskeletal problems within dental students: we aimed to determine the prevalence of these disorders.Design Single centre cross-sectional study.Setting A UK Dental School 2015.Methods Students completed a modified Nordic pain questionnaire.Main outcome measures Self-reported frequency and severity of pain, fitness and coping strategies.Results 63% of 390 respondents were female and 75% aged under 23. Seventy-nine percent experienced pain with 42% experiencing pain for 30 or more days in the past year. Lower back pain was most common (54%) and was most frequently the worst area of pain (48%). Thirty-six percent reported pain lasting at least four hours. The mean 'average pain intensity' VAS score was 3.81/10 (sd = 1.75) and mean 'worst pain intensity' was 5.56 (sd = 2.10). More females reported neck pain (58% versus 37%, P <0.001) and higher 'average pain intensity' (mean 4.02, sd 1.82 versus 3.43 sd 1.55, P = 0.012. Daily stretching was used by 55.7% of respondents, and this positively correlated with 'average' and 'worst pain intensity' (P = 0.096 and P = 0.001) scores. Eighteen percent sought professional help to manage pain.Conclusion Musculoskeletal pain is a problem for dental students. Education in self-care may be helpful; however, assessments of possible interventions are needed.

[Evaluation of the benefit of different complementary exams in the search for a TB diagnosis algorithm for HIV patients put on ART in Niamey, Niger]. / Évaluation du bénéfice apporté par des examens complémentaires pour le diagnostic de la tuberculose chez des patients infectés par le VIH mis sous ARV à Niamey, Niger.

In Niger, the tuberculosis (TB) screening among people living with human immunodeficiency virus (HIV) (PLHIV) is nonsystematic and the use of additional tests is very often limited. The objective of this research is to evaluate the performance and the cost-effectiveness of various paraclinical testing strategies of TB among adult patients with HIV, using available tests in routine for patients cared in Niamey. This is a multicentric prospective intervention study performed in Niamey between 2010 and 2013. TB screening has been sought in newly diagnosed PLHIV, before ART treatment, performing consistently: a sputum examination by MZN (Ziehl-Nielsen staining) and microscopy fluorescence (MIF), chest radiography (CR), and abdominal ultrasound. The performance of these different tests was calculated using sputum culture as a gold standard. The various examinations were then combined in different algorithms. The cost-effectiveness of different algorithms was assessed by calculating the money needed to prevent a patient, put on ART, dying of TB. Between November 2010 and November 2012, 509 PLHIV were included. TB was diagnosed in 78 patients (15.3%), including 35 pulmonary forms, 24 ganglion, and 19 multifocal. The sensitivity of the evaluated algorithms varied between 0.35 and 0.85. The specificity ranged from 0.85 to 0.97. The most costeffective algorithm was the one involving MIF and CR. We recommend implementing a systematic and free direct examination of sputum by MIF and a CR for the detection of TB among newly diagnosed PLHIV in Niger.

Thermolabile phenotype of carnitine palmitoyltransferase II variations as a predisposing factor for influenza-associated encephalopathy.

To assess the etiology of influenza-associated encephalopathy (IAE), a surveillance effort was conducted during 2000-2003 in South-West Japan. All fatal and handicapped patients except one (4/34 patients) exhibited a disorder of mitochondrial beta-oxidation evoked by the inactivated carnitine palmitoyltransferase II (CPT II) with transiently elevated serum acylcarnitine ratios (C(16:0) + C(18:1))/C(2) > 0.09 during high-grade fever. Analyses of genotypes and allele compositions of CPT II revealed a thermolabile phenotype of compound heterozygotes for [1055T > G/F352C] and [1102G > A/V368I], which shows a higher frequency in IAE patients than healthy volunteers (P < 0.025). The thermolabile phenotype of CPT II variations may be a principal genetic background of IAE in Japanese.

Effects of gadolinium chloride (GdCl(3)) on the appearance of macrophage populations and fibrogenesis in thioacetamide-induced rat hepatic lesions.

Macrophages infiltrating injured tissue play an important part in fibrogenesis. To shed light on the functional roles of macrophages, we investigated the appearance of macrophage populations in thioacetamide (TAA)-induced rat hepatic lesions, with or without pretreatment with GdCl(3), a chemical capable of inhibiting Kupffer cell functions. In the GdCl(3)+TAA group rats received a single intraperitoneal injection of GdCl(3) (7.5mg/kg body weight) and, after 24h, a single intravenous injection of TAA (300mg/kg body weight). Rats in the TAA group received TAA only. Rats in both groups were examined on post-TAA injection (PTI) days 3, 5, and 7. In the TAA group, on PTI day 3, when TAA-induced hepatocyte injury was particularly prominent, the number of macrophages peaked, subsequently decreasing until PTI day 7. As compared with the TAA group, the GdCl(3)+TAA group showed significantly decreased numbers of ED1-immunolabelled cells (exudate macrophages) and ED2-immunolabelled cells (Kupffer cells) on PTI days 3, 5, and 7, and OX6-immunolabelled cells (antigen-presenting macrophages) on PTI days 3 and 5. Although less strikingly, the numbers of alpha-smooth muscle actin-positive myofibroblasts and fibrotic areas were decreased in the GdCl(3)+TAA group. By RT-PCR, the expression of TGF-beta1 mRNA was suppressed on PTI days 3 and 7 in the GdCl(3)+TAA group, and the suppressed expression was confirmed in vitro by treating rat macrophage-like cells (HS-P) with 1% GdCl(3). The study showed that GdCl(3) treatment decreased the numbers of macrophages appearing in hepatic lesions and inhibited TGF-beta1 mRNA expression in macrophages. Decreased numbers of macrophages may contribute to improvement of hepatic fibrosis.

Monoclonal antibodies which differentiate high- and low-affinity binding sites of interleukin-2.

Five monoclonal antibodies (L15, L20, L23, L34, and L61) against human recombinant interleukin-2 were tested for their effects on the interleukin-2 bioactivity and binding. Four of these monoclonal antibodies, L15, L20, L34, and L61, which had neutralizing activity, completely blocked interleukin-2 binding to the high-affinity receptor. On the other hand, L23, which had a very weak neutralizing activity, blocked interleukin-2 binding to the low-affinity receptor. These results suggest that there are at least two distinct binding sites on the interleukin-2 molecule; those for the high-affinity receptor and those for the low-affinity receptor. These monoclonal antibodies should be useful tools in the study of the interaction between interleukin-2 and interleukin-2 receptor.

Conditioned media of glial cell lines induce alkaline phosphatase activity in cultured artery endothelial cells. Identification of interleukin-6 as an induction factor.

Conditioned media of human glial cell lines induced alkaline phosphatase activity in cultured calf artery endothelial cells. The maximal alkaline phosphatase activity in the culture was comparable to the level in isolated brain capillary endothelial cells. An induction factor in the conditioned media was purified and identified as interleukin-6 from its amino-terminal sequence, molecular weight, amino acid composition and immunoreactivity. Recombinant interleukin-6 had similar induction activity. Our findings raise the possibility that interleukin-6 induces and modulates alkaline phosphatase activity in endothelial cells during normal development of the blood-brain barrier and under certain pathological conditions.

Familial amyloidotic polyneuropathy diagnosed by cloned human prealbumin cDNA.

A diagnosis of familial amyloidotic polyneuropathy (FAP) can be made by use of restriction endonuclease NsiI, a cloned human prealbumin cDNA and Southern blot procedures. Digests of DNAs from 10 disease-free individuals showed two bands (6.6 kb and 3.2 kb) complementary to a human prealbumin cDNA, whereas digests from 11 individuals with FAP exhibited two additional bands (5.1 kb and 1.5 kb). We interpret these changes in pattern to be the result of a restriction site for NsiI located in the altered codon and associated with the mutant prealbumin gene. All these individuals with FAP were heterozygous for the prealbumin gene, carrying one normal and one mutant gene.

Acetyl-low density lipoprotein receptors on rat mesangial cells.

To elucidate whether mesangial cells have any scavenger functions for modified lipoproteins, surface binding and cholesteryl ester (CE) formation by acetyl-low density lipoproteins (acetyl-LDL) have been studied in cultured rat renal mesangial cells. Specific binding kinetics for acetyl-LDL were observed with Kd = 28.3 micrograms/ml and Bmax = 1.1 ng/micrograms cell protein at 0 degrees C. The fluorescence microscopic finding demonstrated the enhanced uptake of DiI-acetyl-LDL in mesangial cells. Incorporation of [14C]oleate into CE was enhanced to 6-fold by loading 30 micrograms/ml of acetyl-LDL on 10 micrograms/ml of [14C]oleate-bovine serum albumin conjugate as compared with the control without lipoproteins (P < 0.05). The CE formation was completely inhibited by chloroquine. The light microscopic finding demonstrated the increased CE deposition by acetyl-LDL, resulting in foam cell formation. These results indicate biochemically and morphologically that the mesangial cells take up acetyl-LDL by receptor-mediated endocytosis, and that cholesterols in acetyl-LDL are converted to CE, resulting in an increased cellular cholesterol content. In conclusion, mesangial cells may have a scavenger function similar to macrophages.

A fluorescence sandwich ELISA for detecting soluble and cell-associated human interleukin-2.

A fluorescence sandwich ELISA was developed for the quantitation of human interleukin-2 (IL-2) using a combination of two anti-human IL-2 antibodies, L61 and biotinylated anti-IL-2 rabbit IgG. The minimum level of IL-2 detectable by this ELISA was 0.3 pg/ml, indicating that this method is about 100 times more sensitive than the usual colorimetric human IL-2 ELISA. The ELISA specifically detected human IL-2 in cell culture supernatants and serum, and also in extracts from mitogen-stimulated peripheral blood mononuclear cells. All of the IL-2 species detected in the supernatants, sera and cell lysates were shown to have a molecular weight of 15,000, as estimated by size exclusion high performance liquid chromatography.

Ammonium chloride exposure inhibits cytokine-mediated eosinophil survival.

To study human eosinophils, their efficient purification from peripheral blood is crucial. Although a number of purification procedures, including discontinuous Percoll and metrizamide density gradient centrifugation, have been used, it has been difficult to isolate eosinophils from normal donors with consistently high yields and purities. Recently, a new isolation technique called magnetic cell separation system (MACS) was reported. To evaluate this procedure, we isolated eosinophils from human peripheral blood using either MACS or the standard discontinuous Percoll density methods, and compared cellular viability, morphology, and response to degranulation stimuli. MACS gave a higher yield of eosinophils than Percoll density centrifugation; for example, 6.6 +/- 1.1 x 10(6) eosinophils were isolated from 20 ml of blood by MACS compared to 6.4 +/- 2.4 x 10(6) from 120 ml by Percoll density gradient. Further, the purity of eosinophils isolated by MACS was 97.1 +/- 0.5% (X +/- SEM) compared to 77.8 +/- 2.9% with Percoll. As part of the MACS protocol, erythrocytes are lysed with either 155 mM ammonium chloride or hypotonic lysis. With 155 mM ammonium chloride treatment, the eosinophils showed a striking reduction in cytokine mediated survival due to interleukin (IL)-3, IL-5 and granulocyte-macrophage colony-stimulating factor (GM-CSF), marked morphologic abnormalities and a reduced degranulation response. With hypotonic lysis, no differences were observed in survival and morphology between eosinophils purified by MACS and Percoll methods; the degranulation responses to stimuli were essentially the same between the two methods. Taken together, these observations suggest that the exposure of eosinophils to 155 mM ammonium chloride results in cellular damage. Therefore, MACS with hypotonic lysis is a useful technique to isolate eosinophils for biological study.

Neutralizing monoclonal antibodies against recombinant human interleukin-2.

A series of hybridoma cell lines which produce monoclonal antibodies (MAbs) against recombinant human interleukin-2 (rIL-2) have been established by fusion of murine myeloma cell line P3-NS1-1-AG4-1 and spleen cells of BALB/c mice which had been immunized with rIL-2. 48 hybridoma strains were selected by a solid-phase screening method which produced MAbs reacting with IL-2: four MAbs, L-15, L-20, L-34, and L-61, exhibited strong inhibition of the proliferating effect of rIL-2 on IL-2-dependent cell lines, NK7 and CTLL-2. L-61, the most potent MAb among them, also neutralized natural human IL-2, while the other three MAbs were unreactive. All the four MAbs were specific to human IL-2: they did not cross-react with mouse or rat IL-2. These MAbs are expected to be useful tools in the investigation of IL-2 function.

18F-FDG PET detection of colonic adenomas.

UNLABELLED: The adenomatous polyp of the colon is clinically important as a precursor of colonic cancer. The aim of this preliminary study was to evaluate the potential usefulness of (18)F-FDG PET for detecting adenomatous polyps of the colon. METHODS: We performed a retrospective study of 110 subjects who underwent both PET study and total colonoscopy. On nonattenuation-corrected PET images, focal distinct FDG accumulation along the large intestine was considered a positive finding, and the PET results were compared with colonoscopic findings. Histology and adenoma size were determined by polypectomy. RESULTS: Fifty-nine adenomatous polyps, 5-30 mm in size, were found in 30 subjects by total colonoscopy. PET findings were positive for 14 of the 59 adenomas (24%). The positivity rate for PET images rose with the increase in size of the adenomas; it was 90% in adenomas (9/10) that were > or =13 mm. The overall false-positive rate was 5.5% (6/110 subjects). CONCLUSION: Increased glucose metabolism is observed in colonic adenomas, and detectability with PET increases with the increase in adenoma size. Adenomas are premalignant lesions, and it is important to realize that colonic adenomas may be found incidentally during an FDG PET study.

Enhancement of bradykinin-induced prostacyclin synthesis in porcine aortic endothelial cells by pertussis toxin. Possible implication of lipocortin I.

Bradykinin-stimulated prostacyclin synthesis in porcine aortic endothelial cells was enhanced by pretreatment of the cells with pertussis toxin or islet-activating protein (IAP) for 5 hr or longer. Although ADP-ribosylation of a protein with a molecular weight of 41-42 kD in the cell membranes was completed by 3 hr after the addition of IAP into the incubation medium, there was good correlation between enhancement of bradykinin-induced prostacyclin synthesis and ADP-ribosylation of the IAP substrate over a wide range of IAP concentrations. Furthermore, even if IAP was removed from the incubation medium at 3 hr, bradykinin-induced prostaglandin synthesis at 24 hr was still potentiated. Cycloheximide and actinomycin D enhanced bradykinin-induced prostacyclin synthesis and apparently blocked the effect of IAP. Since this result suggested the involvement of an inhibitor protein(s) of prostacyclin synthesis in the IAP effect, we studied the effect of IAP on the level of lipocortin I which is known to inhibit phospholipase A2. Western and Northern blot analyses revealed that IAP decreased the amounts of protein and mRNA of lipocortin I. These results suggest that the enhancement of bradykinin-induced prostacyclin synthesis by IAP is associated with a decrease in the level of lipocortin I.

Treatment of ulcerative colitis with germinated barley foodstuff feeding: a pilot study.

BACKGROUND: Germinated barley foodstuff (GBF) has been shown to attenuate intestinal injury in animal models, largely by increasing luminal short-chain fatty acid production. AIM: To investigate the safety and efficacy of GBF in the treatment of ulcerative colitis (UC). METHODS: Ten patients with active UC received 30 g of GBF daily for 4 weeks in an open-label treatment protocol while the baseline anti-inflammatory therapy was continued. The response to treatment was evaluated clinically and endoscopically. Pre- and post-treatment stool concentrations of short-chain fatty acids were measured by gas-liquid chromatography. RESULTS: Patients showed improvement in their clinical activity index scores, with a significant decrease in the score from 6.9+/-1.4 to 2.8+/-1.5 (mean+/-S.E.M., P < 0.05). The endoscopic index score fell from 6.1+/-2.3 to 3.8+/-2.3 (P < 0.0001). Patients showed an increase in stool butyrate concentrations after GBF treatment (P < 0.05). No side-effects were observed. CONCLUSIONS: Oral GBF therapy may have a place in management of ulcerative colitis, but controlled studies are needed to demonstrate its efficacy in the treatment of this disorder.