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Cytokeratins (CKs) are now considered to be reliable markers for following the development and differentiation of epithelial tissue. We have investigated the pathway of differentiation in human developing junctional epithelium using monoclonal antibodies and two-dimensional gel electrophoresis of microdissected tissue to identify CK 19, CK 16, CK 14, CK 13, CK 6, CK 5, CK 4 in the junctional epithelium (JE) over partially erupted human teeth. The CK profile was similar to that of developing oral epithelia, suggesting that the junctional epithelium in teeth during eruption is of odontogenic origin. The present study used in situ hybridization to determine the distribution of the mRNAs of CKs 19, 16, 13 and 4 in human developing junctional epithelium and to examine the correlation between mRNAs and their encoded proteins. CK 19 mRNA was abundant in the basal cell layers of the primary junctional epithelium (PJE) but less concentrated in the suprabasal layers. CK16, 13 and 4 mRNAs were abundant in the basal cell layers of the PJE. The parabasal cell layers reacted intensely to the cRNA probe complementary to CK16 mRNA, as were the reactions in the suprabasal cell layers of the PJE for the CK 13 and 4 probes. Our results demonstrate that the PJE express the genes encoding for CKs 16 and 4 that have been revealed previously only by electrophoresis. They therefore confirm that the PJE is a well-differentiated stratified epithelium with a complex unique phenotype that produces CKs specific for basal cells (CK 19), CKs associated with hyperproliferation (CK 16), and finally those associated with stratification (CKs 4 and 13). Only synthesis of CK 19 protein and mRNA are strictly parallel. CKs 4 and 13 mRNAs are present in basal and suprasal cells, while their encoded proteins were not, except for CK 13 in suprabasal cell layers of PJE, where the amount of its mRNAs was coincident with the expression of the protein. [ABSTRACT FROM AUTHOR]

This study was undertaken to evaluate the type of tissue obtained after transplantation of a connective tissue graft, from the same palatal donor site, into a non-keratinized oral mucosa in humans. In 6 human volunteers, a thick palatal epithelio-connective tissue graft was excised, split into two thinner grafts (a thin epithelio-connective tissue graft and a connective tissue graft) and transplanted into controlateral areas lacking keratinized gingiva. The biopsies, excised 3 months post-operatively, were examined using routine histology, immunofluorescence techniques with different anticytokeratin antibodies and biochemical techniques with non-equilibrium two-dimensional gel electrophoresis. The results show that the epithelio-connective tissue grafts display the histological and biochemical characteristics of keratinized gingiva, whereas the connective tissue grafts expressed features belonging both to keratinized and non-keratinized gingival tissues, It is concluded that the deep palatal connective tissue does not have the full potential to induce non-keratinized epithelial cells to keratinize and that a gingival or palatal connective tissue graft without its overlying epithelium is not likely to yield genuine attached gingiva. [ABSTRACT FROM AUTHOR]

This study uses cytokeratins (CK) as markers to investigate the phenotype of the junctional epithelium (JE) in partially erupted human teeth. The gingival samples, which were clinically healthy, were carefully dissected from the teeth. Cryostat sections were cut for histological staining, immunofluorescence microscopy and gel electrophoresis. Cytokeratins were extracted after microdissection. The basal and suprabasal epithelial cell markers, cytokeratins 4, 5, 13, 14 and 19 were detected with specific monoclonal antibodies. They showed that the junctional epithelium in erupting teeth has a complex topography. The cytokeratin immunohistochemical profile distinguished between the primary junctional epithelium (CK 5, 14 and 19 in basal and suprabasal cells and CK 13 faintly stained throughout the suprabasal layers) and the adjacent epithelium that had the same cytokeratin profile as the sulcular epithelium (CK 5, 14 and 19 in basal cells and CK 4 and 13 intensively stained in the suprabasal cells). Extraction, two-dimensional electrophoresis and western blotting showed that this transitional JE during eruption also contained CK 6, 16 and perhaps CK 4. Thus, the JE in erupting teeth shows patterns of CK distribution that are very similar to that of developing oral epithelia. [ABSTRACT FROM AUTHOR]

There is scarce information on the current importance of oyster beds as fishing grounds in the United Arab Emirates (UAE). This study aims to understand the socio-economic value of oyster bed fisheries through questionnaire-based surveys with fishers. Of 106 Emirati fishers interviewed, 67 % use oyster beds due to the proximity to shore, better catch quality, and species abundance. Oyster bed fisheries are recreational and commercial, with handline and fish traps the most common used gears. They provide food for local consumption and cash income. All respondents noticed a fish abundance and size decrease throughout the last decade. Fishers suggest establishing marine protected areas and updating fishing regulations to improve fishing stock status. During the Covid-19 pandemic, oyster fisheries increased, highlighting the value of these fishing grounds for food availability. These fisheries support the local economy and heritage, and urgently need management to ensure the protection of these often-overlooked habitats., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

B and T lymphocyte attenuator (BTLA) is a co-inhibitory receptor that is expressed by lymphoid cells and regulates the immune response. Consistent with an inhibitory role for BTLA, the disease is exacerbated in BTLA-deficient lupus mice. We recently demonstrated that the BTLA pathway is altered in CD4 + T cells from lupus patients. In the present work, we aimed at delineating the expression pattern of BTLA on CD4 + T cell subsets suspected to play a key role in lupus pathogenesis, such as circulating follicular helper T cells (cT FH ) and regulatory T cells (Tregs). We did not detect significant ex vivo variations of BTLA expression on total CD4 + T cells (naive and memory), cT FH or T FH subsets between lupus patients and healthy controls. However, we interestingly observed that BTLA expression is significantly increased on activated Tregs, but not resting Tregs, from lupus patients, especially those displaying an active disease. Moreover, it correlates with the diminution of the Tregs frequency observed in these patients. We also showed that both BTLA mRNA and protein expression remain low after TCR stimulation of activated Tregs sorted from healthy donors and evidenced a similar dynamic of BTLA and HVEM expression profile by human Tregs and effector CD4 + T cells upon T cell activation than the one previously described in mice. Finally, we observed that the HVEM/BTLA ratio is significantly lower in Tregs from lupus patients compared to healthy controls, whereas ex vivo effector CD4 + T cells express higher BTLA levels. Our data suggest that an altered expression of BTLA and HVEM could be involved in an impaired regulation of autoreactive T cells in lupus. These results provide a better understanding of the BTLA involvement in lupus pathogenesis and confirm that BTLA should be considered as an interesting target for the development of new therapeutic strategies., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Aubergeon, Sawaf, Felten, Gottenberg, Dumortier and Monneaux.)

Background: There are many laparoscopic techniques for pediatric congenital inguinal hernia repair. Needlescopic surgery was introduced recently in pediatric patients aiming at getting excellent cosmetic outcomes., Purpose: The aim of this study was to describe a novel technique for needlescopic inguinal hernia repair in children., Patients and Methods: Needlescopic division of the hernial sac was carried out on 369 children in 6 pediatric tertiary centers during the period from August 2016 to May 2019. All hernias were repaired by a novel needlescopic procedure that replicates all the steps of the open herniotomy., Results: A total of 369 patients with 410 hernias were included in this study. They were 232 (62.9%) males and 137 (37.1%) females, with a mean age of 3.58±1.26 (range=2 to 8 y) and mean internal inguinal ring diameter was 13.65±3.85 mm (range=8 to 20 mm). The mean operative time was 23.36±4.67 minutes for bilateral and 14.28±2.98 minutes for unilateral cases. All cases were completed without conversion to conventional laparoscopy. All cases were followed up for a mean of 19.6±3.2 months. None of our patients developed recurrence or testicular atrophy and the scars were nearly invisible 3 months postoperatively., Conclusions: Needlescopic pediatric inguinal hernia repair using disconnection of the hernia sac at internal inguinal ring with purse-string suture closure of peritoneum is feasible and safe with no recurrence and with outstanding cosmetic results., Competing Interests: The authors declare no conflicts of interest., (Copyright © 2021 Wolters Kluwer Health, Inc. All rights reserved.)

Background: Laparoscopic Nissen fundoplication is the gold standard antireflux procedure in pediatric age group. Intrathoracic migration of the fundic wrap is a common cause failure, leading to recurrence of gastroesophageal reflux disease (GERD) symptoms. Objectives: To investigate the impact of wrap-crural fixation and minimal esophageal dissection in prevention of wrap transmigration after laparoscopic Nissen fundoplication in children. Methods: Prospective randomized study of 46 pediatric patients with refractory GERD who underwent laparoscopic Nissen fundoplication divided into two equal groups. In Group A, wrap crural fixation was done, whereas in group B no fixation was done. Minimal esophageal dissection with preservation of the phrenoesophageal ligament was done in both groups. Approval of the Ethics Committee of our Faculty was obtained. Results: There was no difference between both groups regarding operative time, intraoperative complications, or length of hospital stay. Two patients in group B without wrap fixation suffered recurrence of GERD symptoms. On contrast study, they both showed intrathoracic wrap migration. One of them was reoperated. Whereas in group A, no recurrence of symptoms and no wrap transmigration were noticed in follow-up. Conclusion: In laparoscopic Nissen fundoplication, with minimal esophageal dissection and preservation of the phrenoesophageal ligament, there is no additional benefit from wrap-crural fixation in prevention of wrap transmigration.

Coinhibitory receptors play an important role in the prevention of autoimmune diseases, such as systemic lupus erythematosus (SLE), by limiting T cell activation. B and T lymphocyte attenuator (BTLA) is an inhibitory receptor, similar to cytotoxic T lymphocyte-associated protein 4 (CTLA-4) and programmed death 1 (PD1), that negatively regulates the immune response. The role of BTLA in the pathogenesis of autoimmune diseases in humans and, more specifically, in SLE is largely unknown. We investigated BTLA expression on various T cell subsets, and we did not observe significant variations of BTLA expression between lupus patients and healthy controls. However, the enhancement of BTLA expression after activation was significantly lower in SLE patients compared with that in healthy controls. Furthermore, we found an impaired capacity of BTLA to inhibit T cell activation in SLE due to a poor BTLA recruitment to the immunological synapse following T cell stimulation. Finally, we demonstrated that defective BTLA function can be corrected by restoring intracellular trafficking and by normalizing the lipid metabolism in lupus CD4+ T cells. Collectively, our results evidence that the BTLA signaling pathway is altered in SLE T cells and highlight the potential of targeting this pathway for the development of new therapeutic strategies in lupus.

In this work, the influence of drug load and physical state of R3040 in self-nanoemulsifying drug delivery systems (SNEDDS) on R3040 absorption in rats was assessed. Furthermore, an in vitro lipolysis model simulating rat conditions (rat lipolysis model) was compared to a human lipolysis model in regard to the prediction of the in vivo data. The formulations were SNEDDS 80%, containing R3040 at 80% of its equilibrium solubility in SNEDDS (S eq ); super-SNEDDS solution with R3040 supersaturated at 200% S eq ; super-SNEDDS suspension containing R3040 at 200% S eq ; Chasing principle (drug-free SNEDDS followed by R3040 aqueous suspension) and R3040 aqueous suspension. The pharmacokinetic profiles of R3040 in SNEDDS 80% and super-SNEDDS solution 200% were superimposed and higher than for super-SNEDDS suspension 200%, Chasing principle and aqueous suspension. Therefore, dosing R3040 dissolved in SNEDDS increased R3040 absorption irrespective of the drug load. While the human lipolysis model could not predict the rank order of absorption of the formulations, the rat lipolysis model predicted the similar absorption of R3040 in SNEDDS 80% and super-SNEDDS solution 200%. Thus, the rat lipolysis model showed to be an important step towards predictive in vitro models for rat studies., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Systemic lupus erythematosus (SLE) is a chronic autoimmune disease characterized by B cell hyperactivity leading to the production of autoantibodies, some of which having a deleterious effect. Reducing autoantibody production thus represents a way of controlling lupus pathogenesis, and a better understanding of the molecular and cellular factors involved in the differentiation of B cells into plasma cells could allow identifying new therapeutic targets. Follicular helper T cells (TFH) represent a distinct subset of CD4(+) T cells specialized in providing help to B cells. They are required for the formation of germinal centers and the generation of long-lived serological memory and, as such, are suspected to play a central role in SLE. Recent advances in the field of TFH biology have allowed the identification of important molecular factors involved in TFH differentiation, regulation, and function. Interestingly, some of these TFH-related molecules have been described to be dysregulated in lupus patients. In the present review, we give an overview of the aberrant expression and/or function of such key players in lupus, and we highlight their potential as therapeutic targets.

Objective: Hirschsprung-associated enterocolitis (HAEC) is one of the most troublesome problems encountered after a pullthrough. We hypothesized that prophylactic administration of probiotics after a pullthrough procedure would decrease the incidence of HAEC., Study Design: A prospective, double-blind, placebo-controlled, randomized trial was conducted at 2 children's hospitals. Infants undergoing pullthrough were randomized to probiotic or placebo for a period of 3 months post-pullthrough. Primary outcome was incidence of post-operative HAEC. Other outcomes included severity of HAEC by clinical grade, number of HAEC episodes and extent of aganglionosis. Pearson Chi Square analysis, as well as logistic regression, was used for statistical analysis., Results: Sixty-two patients were recruited (Sites: A=40; B=22). One was lost to follow up and one immediate post-op death was not included in final analysis. Probiotics were administered to 32 patients. Distribution of placebo/probiotics was equal between sites (P=0.858). Mean age at pullthrough was 6.5 ± 8.1(± SD) months. The incidence of HAEC was 28.3%. The incidence of HAEC was not statistically different between probiotic and placebo study groups., Conclusions: Incidence of HAEC was not reduced with prophylactic probiotics. Future studies are needed to better determine the etiology and possible ways of preventing this complex condition., (Copyright © 2013 Elsevier Inc. All rights reserved.)

Background and Study Aims: Despite the growing understanding of the involvement of protooncogenes and tumour suppressor genes in the oncogenesis of CRC, the exact biological and molecular mechanisms underpinning this process remain poorly understood. The signal transducer and activator of transcription (STAT3) has been implicated in the regulation of growth and malignant transformation. Accumulating evidences have come to indicate that abnormalities in the Janus kinase (JAK)/STAT pathway are involved in oncogenesis of several cancers. The aim of this study was to investigate the expression of JAK3 and STAT3 in both normal and activated forms by immunohistochemistry in adenomas of the colon, ulcerative colitis and CRC compared to normal colonic mucosa., Patients and Methods: Tissues from 30 cases with primary CRC and seven cases with ulcerative colitis (UC), removed by colectomy, were included. In addition, tissues from 10 colonic adenomas, 15 CRC and eight cases with UC, obtained by endoscopic biopsies, were examined histopathologically. Immunohistochemical evaluation of STAT3, p-STAT3, JAK3 and p-JAK3 expression in tissue sections was completed. Statistical analysis and correlation of data were then performed., Results: Normal colonic mucosa showed expression of STAT3 only. Immunoreactivity of p-JAK3 increased significantly (p<0.05) and correlated with the degree of dysplasia in colonic adenomas. Immunoreactivity of p-STAT3 increased significantly (p<0.05) and correlated with the degree of dysplasia in cases with UC. In CRC a significant positive correlation was found between p-STAT3 expression and grading, STAT3, JAK3 and p-JAK3 and TNM or Dukes' staging, and p-STAT3 and nodal status excluding distant metastasis (p<0.05)., Conclusion: JAK3 and STAT3, and particularly their activated forms, were found to correlate significantly with the degree of dysplasia in adenomas and UC, indicating their potential role in colorectal carcinogenesis. They also correlate with anaplasia and invasion, suggesting a definitive role in progression of CRC., (Copyright © 2009 Arab Journal of Gastroenterology. Published by Elsevier Ltd. All rights reserved.)

Background: Recent work indicates that mechanical force induces small-bowel growth, although methods reported do not have direct clinical application. We report a clinically feasible technique of enterogenesis and describe intestinal function in this model., Methods: Using a pig model (n = 11), we stretched isolated small intestinal segments mechanically for 7 days in vivo with an intraluminal device. Control segments were not stretched. Morphology, histology, and epithelial proliferation were assessed. Absorption and epithelial barrier function were examined in an Ussing chamber., Results: Stretch segments were significantly longer than Control segments and had nearly 2-fold greater surface area (P < .001). Mucosal thickness was much greater in Stretch than Control segments (772 +/- 134 vs. 647 +/- 75 microm, P = .02). Although villus height was reduced in Stretch and Control segments (353 +/- 76 vs. 324 +/- 76 microm, P = .6) versus native jejunum (522 +/- 87, P < .0005), crypt depth was increased dramatically in Stretch (450 +/- 95 microm) versus Control segments (341 +/- 64, P = .005). This observation was accompanied by a 2-fold increase in cellular proliferation (26.3 +/- 3.8 vs 12.1 +/- 6.6 % bromodeoxyuridine+, P < .05). Barrier function was intact ([3H]-mannitol permeation, 0.16 +/- 0.08%, vs native jejunum, 0.17 +/- 0.08%, P = .81). Glucose-mediated sodium transport was similar in Stretch versus native jejunum segments (60.0 +/- 23.5 vs 82.3 +/- 47.3 microA/cm2, P = .31), as was carbachol-induced chloride transport (82.4 +/- 72.2 vs 57.2 +/- 33.4 microA/cm2, P = .54) and alanine absorption (16.46 +/- 12.94 vs 23.53 +/- 21.31 microA/cm2, P = .53)., Conclusions: Mechanical stretching induces small intestinal growth, while maintaining function. Epithelial architecture does change, such that a decrease in villus height is offset by a marked increase in crypt depth and a 2-fold increase in epithelial proliferation. Epithelial barrier and absorptive functions remain intact. The device described may have direct clinical applicability.

To assess the attitude and knowledge of physicians and patients towards psychiatry, we asked 115 referring doctors and 188 referred patients to complete questionnaires. We examined the results along with the referral rates to try to identify factors that may affect a consultation-liaison psychiatry service. Generally, knowledge was poor and attitudes towards psychiatry negative in both groups. This negatively influenced the referral rates and reflected the lack of integration of psychiatry and medicine at the training level. This is an indication that psychiatrists need to work in collaboration with hospital doctors to integrate psychiatry into medicine at all levels and emphasizes the priority of education of hospital staff, patients and the community in consultation-liaison psychiatry.

Consultation-liaison psychiatry has emerged as an important sub-specialty in the general hospital setting during recent years as a result of psychiatric acute wards moving into these hospitals. This has inspired the need for better structured research to establish its relevance and effectiveness. We, therefore, carried out a prospective cohort study at King Fahad General Hospital. We report the interaction of sociodemographic, clinical and diagnostic factors, time lag of referral and diagnostic ability of referring physicians. A total of 206 patients were referred over a period of 6 months. Sensitivity and specificity of the diagnostic skills of the referring doctors were found to be generally poor, particularly for anxiety.

With the advent of the new sophisticated carbondioxide (CO2) laser, effective treatment of facial rhytides and pigmented lesions of the photodamaged skin has been gaining popularity. Partial and full facial skin resurfacings have been reported. However, resurfacing of the neck has been ignored or discouraged. The objective of this study is to assess the feasibility, benefits, and risks of laser resurfacing of the neck, combined with the additional resurfacing of the whole face. A total of 48 patients underwent the procedure with a high-power and short-exposure time CO2 laser (ultrapulse). Patients were evaluated and classified preoperatively. The great majority of patients had local anesthesia (topical and infiltrative) with minimal sedation. A computer pattern generator was used in all cases but one. In this paper, preoperative preparation, postoperative care, laser resurfacing techniques, and depth guidelines are discussed. Incorporating the neck with the facial resurfacing has given very satisfactory results, with minimal drawbacks.

Cytokeratins (CKs) are now considered to be reliable markers for following the development and differentiation of epithelial tissue. We have investigated the pathway of differentiation in human developing junctional epithelium using monoclonal antibodies and two-dimensional gel electrophoresis of microdissected tissue to identify CK 19, CK 16, CK 14, CK 13, CK 6, CK 5, CK 4 in the junctional epithelium (JE) over partially erupted human teeth. The CK profile was similar to that of developing oral epithelia, suggesting that the junctional epithelium in teeth during eruption is of odontogenic origin. The present study used in situ hybridization to determine the distribution of the mRNAs of CKs 19, 16, 13 and 4 in human developing junctional epithelium and to examine the correlation between mRNAs and their encoded proteins. CK 19 mRNA was abundant in the basal cell layers of the primary junctional epithelium (PJE) but less concentrated in the suprabasal layers. CK16, 13 and 4 mRNAs were abundant in the basal cell layers of the PJE. The parabasal cell layers reacted intensely to the cRNA probe complementary to CK16 mRNA, as were the reactions in the suprabasal cell layers of the PJE for the CK 13 and 4 probes. Our results demonstrate that the PJE express the genes encoding for CKs 16 and 4 that have been revealed previously only by electrophoresis. They therefore confirm that the PJE is a well-differentiated stratified epithelium with a complex unique phenotype that produces CKs specific for basal cells (CK 19), CKs associated with hyperproliferation (CK 16), and finally those associated with stratification (CKs 4 and 13). Only synthesis of CK 19 protein and mRNA are strictly parallel. CKs 4 and 13 mRNAs are present in basal and suprasal cells, while their encoded proteins were not, except for CK 13 in suprabasal cell layers of PJE, where the amount of its mRNAs was coincident with the expression of the protein.

The effects of different intracanal medicaments and zinc oxide-eugenol based root canal sealers on an experimentally induced apical periodontitis were studied histologically in mesial roots of lower molars of Wistar rats. After root canal instrumentation each canal was filled either with an intracanal medicament twice for 3 days each time or with a sealer for 21 days. The intracanal medicament chlorophenol caused periapical tissue damage. P-chloroxylenol-camphor lead to periapical improvement as did 5% sodium-hypochlorite. 12% sodium-hypochlorite gave tissue damage. After intracanal medication with an aqueous suspension of calcium hydroxide periapical repair was clearly visible. A combination of a corticoid and an antibiotic appeared to induce damage of the periapical tissues. The root canal sealers Endomethasone and N2 essentially impaired periapical repair. The results were acceptable for Aptal-Zink-Harz root canal fillings after 21 days as well as after 56 days.

This study uses cytokeratins (CK) as markers to investigate the phenotype of the junctional epithelium (JE) in partially erupted human teeth. The gingival samples, which were clinically healthy, were carefully dissected from the teeth. Cryostat sections were cut for histological staining, immunofluorescence microscopy and gel electrophoresis. Cytokeratins were extracted after microdissection. The basal and suprabasal epithelial cell markers, cytokeratins 4, 5, 13, 14 and 19 were detected with specific monoclonal antibodies. They showed that the junctional epithelium in erupting teeth has a complex topography. The cytokeratin immunohistochemical profile distinguished between the primary junctional epithelium (CK 5, 14 and 19 in basal and suprabasal cells and CK 13 faintly stained throughout the suprabasal layers) and the adjacent epithelium that had the same cytokeratin profile as the sulcular epithelium (CK 5, 14 and 19 in basal cells and CK 4 and 13 intensively stained in the suprabasal cells). Extraction, two-dimensional electrophoresis and western blotting showed that this transitional JE during eruption also contained CK 6, 16 and perhaps CK 4. Thus, the JE in erupting teeth shows patterns of CK distribution that are very similar to that of developing oral epithelia.

Subepithelial palatal connective tissue grafts, separated from the epithelium either chemically (n = 5) or surgically (n = 2) were inserted in patients presenting with gingival recession. Biopsies at the grafted tissue and a portion of non-keratinized mucosa were taken 12 months later. Histology showed keratinization of the newly formed epithelium, and interestingly a deep projection of epithelium into the connective tissue in almost all biopsies, sometimes with an enlargement and a cyst-like space. We conclude that chemical separation of epithelium and connective tissue is clinically feasible for connective tissue grafts and that the subepithelial connective tissue grafting technique should be modified to avoid this projection of epithelium.

Epithelial cell intermediate filaments, or cytokeratins, are excellent markers for cell differentiation. During embryogenesis, cytokeratins specific of a stage of differentiation step always become detectable before corresponding morphologic changes: for instance, cytokeratins 5 and 14 are found around the eight week, shortly before stratification of the epithelium occurs, and cytokeratins 1 and 10 are produced before morphologic evidence of keratinization becomes detectable. Among potential diagnostic applications, analysis of cytokeratin patterns of epidermal cells desquamated in the amniotic fluid may provide earlier and less invasive diagnosis than fetoscopic biopsies. Similarly, a review of cytokeratins expressed in a variety of epithelial diseases (involving the epidermis, digestive tract, respiratory tract, urogenital tract, or breast) demonstrated persistence of the original tissue pattern in some instances (this was the case for the majority of simple epithelia) but not in others (complex epithelia). This suggests that cytokeratins may prove valuable as markers for specific tumor stages or types and may provide earlier information than morphologic studies.

Intermediate filaments, the most stable of cytoskeleton components, are extremely diverse and usually correlate with the histological subtype since in nearly all cell types a single type of intermediate filament (IF) is found. The cytokeratins, which are specific of epithelia, are the largest and most diverse class of intermediate filaments. Twenty different cytokeratin polypeptides have been identified in humans and separated on the basis of isoelectrical pH and apparent molecular weight using two-dimensional electrophoresis. These data have been used to establish a cytokeratin catalogue which currently serves as a reference [43, 48]. The number of cytokeratin polypeptides expressed ranges from 2 to 5 for each epithelial cell and from 2 to 10 for each epithelium and even of each cell layer within a given epithelium. A broad spectrum of anticytokeratin antibodies with subgroup or single polypeptide specificity is currently available. The distribution of cytokeratins in normal epithelia is reviewed herein and commercially available anti-cytokeratin antibodies are listed.

Low density gingival epithelial cells were cultured on the side of glass slides facing rat's tail collagen lattices. Under these conditions and in the presence of physiological level of calcium, colony formation was enhanced and stratification was slowed down. The strong attachment of the cells to glass slides permitted immunocytochemical examination of cytokeratin (CK) expression and their organization within individual cells during the different stages of epithelial maturation. The present results showed that during the stage of cell migration and colony formation, the cells express the same set of cytokeratins (basal cell marker 14, simple epithelial markers 8, 18 and 19, and marker of hyperproliferation 16) which forms a well-defined network of organized filaments. At the stratification stage, the filament network became dense by the additional expression of the markers of differentiation in non-keratinized stratified epithelia (CK 4 and 13). These appeared once individual cells started to overlap the basal cells, a period during which the cell-temporarily changed morphology. Whilst the suprabasal cells exhibited dense filament network labelled for CK 4 and 13, the density of labelled filaments for CK 14, 8 and 18 was much lower, indicating that these cells contained newly-formed filaments lacking the basal and simple epithelial keratins. The simple epithelial cytokeratins became weakly labelled in older cultures. The uncoupling of paired expression of cytokeratins 4 and 13 was observed in non-colony forming aged cells. This provides an example of altered program of cytokeratin expression during epithelial maturation.

This article proposes a classification of oral epithelial and summarizes recent literature of cytokeratin expression in the oral cavity. The oral epithelial are subdivided into two major groups: superficial and deep epithelia. Epithelial lining the oral cavity, i.e., superficial, are essentially stratified squamous epithelia, with the exception of taste buds. The epithelium covering the dorsal tongue is a combination of keratinized and non-keratinized epithelia. Deep epithelia are of two kinds, odontogenic and glandular epithelia. This study provides a classification of the cytokeratins expressed in the oral cavity in healthy and pathological situations based on published data and our own studies. The profiles of these polypeptides in different oral epithelia should provide information that may be used in various disciplines of oral medicine.

The characterization of cytokeratin (CK) in adult oral mucosa and developing teeth have been well documented in human. Cytokeratin distribution in developing oral mucosa has not yet been described. The aim of this study was to identify the expression of CK in human fetal tongue (week 10 to week 23) and to correlate the results with morphological maturation. Simple epithelial CK are expressed in all cell layers during the early stages, essentially in peridermal cells. From the 14th week, CK 18 is present only in the taste buds, making this polypeptide a reliable marker for this sensory organ. CK 4 and 13 are expressed from the 10th to the 23rd week by both ventral and dorsal lingual epithelia. Terminal differentiation keratins (CK 1, 2 and 10-11) can only be detected immunohistochemically at the 14th week in some cells on the external surface of some papillae. The number of these papillae and positive cells increase at the 19th and 23rd weeks. The terminal differentiation markers are expressed several weeks earlier than the formation of a well-distinguished keratinized layer.

Previous studies have related preoperative status and severity of disease to the outcome of coronary artery bypass surgery. Although increased perfusion and clamp times increase the risk of cardiac surgical procedures, the importance of these factors in relation to the patient's preoperative condition and the severity of disease has not previously been determined. In this study of 1078 patients, we examined the correlation between the patient's preoperative condition, the severity of coronary disease, and duration of perfusion and clamp time, and the type of oxygenator used with the mortality and morbidity associated with coronary artery bypass grafting. One-way analysis of variance and multiple correlation analysis showed that perfusion time, clamp time and nonclamp perfusion time correlated with mortality, perioperative infarction, the use of intra-aortic balloon pump, stroke, renal failure, pulmonary failure, infection, and leg wound complications (p less than 0.05). Perfusion time, clamp time and nonclamp perfusion time did not correlate with postoperative bleeding or sternal wound complications. Nonclamp perfusion time correlated more strongly than any other factor with mortality, perioperative infarction, the use of intra-aortic balloon pump, renal failure, pulmonary failure and infection (p less than 0.05). Clamp time correlated more than any other factor with the development of leg-wound complications (p less than 0.05). The use of a bubble rather than a membrane oxygenator was significantly related to mortality, stroke, infection and leg wound complications by one-way analysis of variance (p less than 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)

In a three-dimensional culture model, oral epithelial differentiation was investigated ultrastructurally and biochemically for cytokeratin expression. Epithelia from the hard palate, gingiva and alveolar mucosa grown on freely floating collagen lattices populated with fibroblasts from homotypic origins, and fed with medium containing 10% delipidized fetal calf serum for 21 days before analysis, stratified and differentiated to basal cuboidal cells, polyhydral spinous cells and elongated superficial cells. The epithelium of palatal origin had non-nucleated superficial cells resembling orthokeratinized cells. The upper spinous cells had keratohyalin-like granules. The corresponding cells of gingival and alveolar mucosal origins retained their nuclei and had smaller numbers of keratohyalin-like granules. Basal cell keratins (CK 5 and 14) and those of hyperproliferation (CK 6 and 16) were consistently found in all epithelia. Furthermore, simple epithelial keratins (CK 18 and 19) were variably expressed by cells from different oral origins. In epithelial cells from the alveolar mucosa, CK 13 and 19 formed major bands, which correlates with their expression in vivo. In contrast, these polypeptides were either absent or formed minor bands in extracts of gingival and hard palatal cells. Although in small quantities, keratins of terminal differentiation (CK 1, 2, 10 and 11) were detected in gels prepared from palatal epithelia. This expression correlates with the higher morphological differentiation of these cells in this model. The model is of interest for studies of epithelial differentiation, as the differentiation markers of keratinized epithelia (CK 1 and 10) were expressed by cells from palatal origin, and those of non-keratinized epithelia (CK 4, 13 and 19) were prominent in cells from alveolar mucosal origin.

Cytokeratin expression by the developing human enamel organ between the 10th and the 23rd gestational week was studied by indirect immunofluorescence microscopy technique using a panel of 15 monoclonal antibodies. The results showed that five antibodies (RKSE 60, Kk 8-60, EE 21-6, 6B10 and 1 C-7) were never reactive, that five antibodies (RCK 102, 42.39.13.1, Ks 19 and Pan 1-8) were always positive and that five antibodies (KB 37, RPN 11-62, Ks 13.1, Ks 8-12 and Ks 18.174) obtained or increased their positivity between weeks 12 and 13. It was concluded that a switch in cytokeratin expression occurred around the 12th-13th weeks. No further important change could be noticed after this period. So it is suggested that final cell differentiation was initiated at weeks 12-13.

A five-centre double-blind crossover trial of two two-week periods using diclofenac and indomethacin showed that both drug groups (51 patients) with rheumatoid arthritis responded similarly in relation to pain scores and morning stiffness. It was noted that the response was better in inpatients than in outpatients, despite differences in disease severity. In the osteoarthritis trial (58 patients) it was shown that neither drug significantly reduced resting pain, although both drugs were significantly better in reducing pain on movement; however, patient preference was for diclofenac. Three patients treated with indomethacin withdrew owing to side-effects, compared with one on diclofenac. A slight but significant decrease in haemoglobin levels was observed in both treatment groups with osteoarthritis, but this did not appear to be symptom-related.

In man, cytokeratin constitutes a family of 19 polypeptides that show different but distinct distribution patterns in the various epithelia. Changes in these patterns may occur during epithelial development and differentiation. The cytokeratin patterns in the oral mucosa of the miniature pig, an animal used in studies of wound healing, were investigated. Surgical biopsies were obtained from the gingiva, hard palate and alveolar mucosa of both man and pig. The cytokeratins were analysed by immunofluorescence, two-dimensional gel electrophoresis and by immunoblotting. Nine monoclonal antibodies were used to identify the different cytokeratin polypeptides in cryostat sections. Two-dimensional gel electrophoresis showed that pig oral mucosa contains at least 10 different polypeptides, five of the acidic type I and five of the basic type II cytokeratins. These were different from the human cytokeratin polypeptides and accordingly were designated P1-P10, according to their molecular weight and isoelectric mobility. Their molecular weight varied between 48 and 69 kdalton and the pHi varied between 5 and 7.3. Immunoblotting showed the monoclonal antibody Ks 13.1 (anticytokeratins Nos 13 and 14) to cross-react with the pig polypeptides P10 and P8. Immunolocalization showed that all the antibodies cross-reacted with the pig tissue except Ks 19.1 (anticytokeratin No. 19). It was possible to differentiate between pig alveolar mucosa, which expressed only P3, P4, P5, P8 and P10, and the gingival and hard palatal mucosae, which expressed all 10 polypeptides except P5. This distinction was made by antibody 6B10 (anticytokeratin No. 4), which reacted only with alveolar mucosa; antibody Ks 13.1, which strongly reacted with uncornified mucosa but weakly with cornified mucosa (gingiva and palate); and any of RKSE60, Kk 8.60 or EE21.6 (anticytokeratin No. 10, anticytokeratins Nos 10 and 11 and anticytokeratins Nos 1, 2, 10 and 11, respectively), which reacted strongly with cornified mucosa but weakly, if at all, with uncornified mucosa. These findings provide a baseline for studies on epithelial differentiation in the miniature pig such as in wound healing.