Adult human cytokeratin 19-positive cells reexpress insulin promoter factor 1 in vitro: further evidence for pluripotent pancreatic stem cells in humans.

Human pancreatic cells with a typical ductal phenotype and potential to proliferate can be obtained in vitro, but the differentiation capacity of these putative human pancreatic stem cells remains to be documented. We investigated the protein and mRNA expression of insulin promoter factor 1 (IPF-1) (or pancreas/duodenal homeobox 1), a transcription factor critical for pancreatic development and endocrine cell neogenesis, in human pancreatic ductal cells derived from cultured exocrine tissue. In vitro, exocrine cells rapidly adhered (within 12 h) and were de-/transdifferentiated to ductal cells after 3 days with a dramatic loss of amylase protein (n = 4, 92 +/- 3.3%, P < 0.05 vs. day 1) and a simultaneous increase of ductal cytokeratin 19 protein (n = 4, 3.4-fold on day 3 and 7-fold on day 9, P < 0.05 vs. day 1). IPF-1 protein and mRNA levels were low to undetectable in exocrine preparations before culture. After 2 days of culture, a 3.2-fold increase in IPF-1 protein was observed, corresponding to the characteristic 46-kDa protein in Western blots. Reverse transcriptase-polymerase chain reaction confirmed a 10.5-fold increase in IPF-1 mRNA levels after 3 days of culture (n = 5, P < 0.001 vs. day 1). Double immunocytochemistry showed direct evidence that IPF-1 appeared during culture in these exocrine-derived ductal cells (cytokeratin 7-positive) and was not merely in contaminating endocrine cells (chromogranin A-positive). In conclusion, we describe herein the first converging evidence on both the molecular and protein level that human cells with a typical ductal phenotype derived ex vivo from pancreatic exocrine tissue (obtained from healthy donors) can reexpress IPF-1 in culture, suggesting their pancreatic precursor/stem cell potential.

Identification and purification of functional human beta-cells by a new specific zinc-fluorescent probe.

Pancreatic beta-cells contain large amounts of zinc. We took advantage of this to try to localize, quantify, and isolate insulin-producing cells from islet preparations. Our study was designed to identify a non-toxic zinc-sensitive fluorescent probe able to selectively label labile zinc in viable beta-cells and to exhibit excitation and emission wavelengths in the visible spectrum, making this technique exploitable by most instruments. We tested Newport Green, a probe excitable at 485 nm with a dissociation constant in the micromolar range corresponding to a low affinity for zinc. The loading of the lipophilic esterified form of Newport Green was easy, rapid, specific, and non-toxic to cells. Confocal microscopy highlighted an intense fluorescence associated with secretory granules. Regression analyses showed a good relationship between zinc fluorescence and islet number (r = 0.98) and between zinc fluorescence and insulin content (r = 0.81). The determination of Zn fluorescence per DNA enabled us to assess the quality of the different islet preparations intended for islet allografting in terms of both purity and viability. Cell sorting of dissociated Newport Green-labeled cells resulted in a clear separation of beta-cells, as judged by insulin content per DNA and immunocytochemical analysis. This zinc probe, the first able to specifically label living cells in the visible spectrum, appears very promising for beta-cell experimentation, both clinically and for basic research.

Beneficial effect of 1,25 dihydroxyvitamin D3 on cytokine-treated human pancreatic islets.

We examined whether 1,25 dihydroxyvitamin D(3) (1,25 D(3)), the active form of vitamin D involved in the regulation of the immune system, may also protect human pancreatic islet cells from destruction induced by cytokines. In this study, we specifically investigated the effect of 1,25 D(3) on oxidative stress and major histocompatibility complex (MHC) induction, both implicated in cytokine-induced islet cell dysfunction and destruction. We also investigated the effects of 1,25 D(3) on interleukin (IL)-6, a pleiotropic cytokine implicated in the pathogenesis of immunoinflammatory disorders. Human pancreatic islets, isolated from heart-beating donors, were treated with a combination of three cytokines, IL-1beta+tumor necrosis factor alpha+interferon gamma, in the presence or absence of vitamin D, and compared with with untreated control cells. Metabolic activity was assessed by cell viability and insulin content. Oxidative stress was estimated by heat shock protein 70 (hsp70) expression, cell manganese superoxide dismutase (MnSOD) activity and nitrite release, a reflexion of nitric oxide (NO) synthesis. Variation of immunogenicity of islet preparations was determined by analysis of the MHC class I and class II transcripts. Inflammatory status was evaluated by IL-6 production. After 48 h of contact with cytokines, insulin content was significantly decreased by 40% but cell viability was not altered. MHC expression significantly increased six- to sevenfold as well as NO and IL-6 release (two- to threefold enhancement). MnSOD activity was not significantly induced and hsp70 expression was not affected by the combination of cytokines. The addition of 1,25 D(3) significantly reduced nitrite release, IL-6 production and MHC class I expression which then became not significantly different from controls. These results suggest that the effect of 1,25 D(3) in human pancreatic islets cells may be a reduction of the vulnerability of cells to cytotoxic T lymphocytes and a reduction of cytotoxic challenge. Hence, 1,25 D(3) might play a role in the prevention of type 1 diabetes and islet allograft rejection.

Rapid detection of enterovirus in clinical specimens using PCR and microwell capture hybridization assay.

A rapid detection method of enteroviral RNA in clinical samples using PCR and a microwell capture hybridization assay is described. PCR products were labelled directly by digoxigenin-dUTP during the amplification step. The labelled amplicons were hybridized with a biotinylated oligo-probe and captured on commercially available test microwells coated with streptavidin. The hybridized amplicons labelled with digoxigenin were detected using anti-digoxigenin Fab fragments conjugated to peroxidase and colorimetric reaction automatically measured. This method detected as few as 0.01 PFU/100 microl of biological sample with a result obtained within 8 h. Using this method, we were able to detect enteroviral RNA in 23 of 35 clinical specimens from 16 of 17 patients with suspected acute or chronic enteroviral infection. The samples included cerebrospinal fluid, broncho-pulmonary lavage, pericardial effusion, throat swabs, stools, sera, muscular and myocardial biopsies. In contrast, virus was isolated in cell culture in only 8 of 28 clinical specimens from 6 of the 17 patients. This easy-to-perform assay has useful potential in the rapid detection of enterovirus in acute or chronic infection. This methodology could be used for a rapid qualitative detection of other RNA viruses.

Association of human herpesvirus 6 infection with drug reaction with eosinophilia and systemic symptoms.

BACKGROUND: There is a current debate regarding the association of human herpesvirus 6 (HHV-6) infection and drug reaction with eosinophilia and systemic symptoms (DRESS). METHODS: Seven consecutive patients hospitalized with DRESS were enrolled in a prospective study to evaluate evidence of active HHV-6 infection. OBSERVATIONS: The imputable drugs were carbamazepine (5 patients), ibuprofen (1 patient), and sulfasalazine (1 patient). All patients were seropositive for anti-HHV-6 IgG antibodies. Anti-HHV-6 IgM antibodies were detected in 4 of the 7 patients with a seroconversion in 2 patients. Neither anti-cytomegalovirus nor anti-Epstein-Barr virus early antigen IgM antibody was detected. Human herpesvirus 6 genome was not detected by polymerase chain reaction in the first serum sample of all patients. It was weakly detected in skin lesions in the last patient tested by polymerase chain reaction but was not found in uninvolved skin. CONCLUSIONS: The results suggest an association between HHV-6 active infection (primo-infection or reactivation) and severe DRESS. Absence of anti-cytomegalovirus or anti-Epstein-Barr virus early antigen IgM antibodies argues against a nonspecific viral reactivation. Human herpesvirus 6 infection may play a role in the development of DRESS in susceptible patients. Some drugs with reactive metabolites could favor reactivation and propagation of HHV-6.

A comparison of the efficacy and safety of lymecycline and minocycline in patients with moderately severe acne vulgaris.

A multicentre, randomised, double-blind and double-dummy study was conducted to compare the efficacy and safety of lymecycline (n = 71) with that of minocycline (n = 73) in 144 patients with moderately severe acne vulgaris. Patients with an acne score of 1-5 on the Leeds scale received oral lymecycline, 300 mg/day for 2 weeks, then 150 mg/day for 10 weeks or oral minocycline, 100 mg/day for 2 weeks then 100 mg every other day for 10 weeks. Inflammatory, non-inflammatory and total lesion counts were determined at baseline (week 0) and after 4, 8 and 12 weeks' treatment, and global efficacy and safety assessments were made by the patient and investigator at the end of the study. Both treatments were equally effective at reducing differential lesion counts and improving acne condition and severity, with no significant differences between treatments. Inflammatory lesions were reduced by 50.6% and 52.2% with lymecycline and minocycline, respectively, and non-inflammatory lesions by 40.6% and 32.2%. Acne severity was reduced by 42.4% with lymecycline and by 47.9% with minocycline. A total of 4.3% of lymecycline recipients and 4.1% of minocycline recipients experienced treatment-related adverse events, the majority of which were mild in nature. Lymecycline was as effective as minocycline for the treatment of moderately severe acne vulgaris. Both treatments were well tolerated, although there were slightly fewer adverse gastrointestinal and dermatological effects with lymecycline.

Global improvement of systemic scleroderma under long-term administration of octreotide.

Octreotide has proven to be effective for the treatment of intestinal dysmotility in patients with scleroderma in short-term administration. We report a global improvement of scleroderma manifestations under long-term administration of octreotide. A 53-year-old black woman was diagnosed with a four-year history of progressive and severe systemic scleroderma, with diffuse skin sclerosis, myositic involvement, impaired carbon monoxide transfer factor (57% of the predicted normal value and severe digestive involvement with pseudo-obstruction and bacterial overgrowth into the intestinal lumen). After one month of octreotide (75 mug/d), oral feeding was restarted and weight gain of 6.5 kg was achieved. After 8 months of treatment, normal weight was obtained and skin induration was spectacularly reduced and pigmentation returned to a normal state. Dyspnea disappeared and physical activity was quite normal. Octreotide effects on intestinal transit are unclear and may be secondary to immunomodulation or neurotransmission effects. Extradigestive effects of octreotide in scleroderma have not been studied. This report suggests that long-term administration of octreotide may be beneficial in the treatment of patients with systemic scleroderma. Long-term trials are required to confirm these preliminary results.

Bullous pemphigoid in a leg affected with hemiparesia: a possible relation of neurological diseases with bullous pemphigoid?

We report a typical case of bullous pemphigoid (BP) associated with a neurological disorder and study a possible link between neurological disorders and BP. An 84-year-old hemiplegic woman presented with unilateral BP on the hemiparetic side. BP was confirmed by histological and immunofluorescence data. The medical records of the previous 46 consecutive patients with BP were retrospectively analyzed (average age: 79; median age: 85). Thirty of the 46 patients with BP had neurological disorders. These disorders included dementia, epilepsy, multiple sclerosis, cerebral stroke, Parkinson's disease, gonadotropic adenoma, trembling, dyskinesia, lumbar spinal stenosis. In a control group of the 46 consecutive oldest patients (older than 71; average age: 82,5; median age: 80) with another skin disease referred during the previous two-year-period to our one-day-unit only, 13 patients had a neurological disorder. This study demonstrates that there is a high prevalence of neurological disorders in patients with BP (p = 0.0004). A prospective case control study with neurological examination and psychometrical evaluation is warranted to confirm these data. We speculate that neuroautoimmunity associated with the aging process or neurological disorders may be involved in pemphigoid development via an autoimmune response against dystonin which shares homology with bullous pemphigoid antigen 1. Bullous pemphigoid could be considered to be a marker of neurological disorder.

[Lympho-epithelial skin tumor. Case report with review of the literature and nosologic discussion]. / Tumeur lympho-épithéliale cutanée. A propos d'un cas avec revue de la littérature et discussion nosologique.

The clinical and histological findings of a lymphoepithelial tumor of the skin are presented and compared to the 11 cases already published in the literature. This review permits to point out the main characteristics of this recently described tumor. In 10 of the 11 cases the tumor is located on the face. The diagnosis of basal cell carcinoma is most often evoked. The mean age is 40 and 9 of the 11 cases are women. The tumor histologically presented as an epithelial proliferation of basaloid cells with peculiar features: cyst-like cavities infiltrated by mononuclear cells, histiocytic and lymphocytic cells; marks of pilosebaceous differentiation: curling of epithelial cells, areas of keratinisation, large clear cells of sebaceous aspect. The origin of this tumor is discussed and the hypothesis are oriented towards a rare histological form of basal cells carcinoma or a hair adnexial tumor. New cases would provide more informations to specify the nosological place of this tumor.

[Epithelioma cuniculatum. Apropos of a case with review of the literature]. / L'epithélioma cuniculatum. A propos d'un cas, avec revue de la littérature.

A case of Epithelioma cuniculatum arising on a leg ulcer in a 85 years old lady is described. 102 cases have been reported before, mainly located on the sole of the foot: this rare tumor may be recognized by the gross appearance of a slow-growing exophytic cauliflower growth, exuding foul smelling material from numerous sinuses, which eventually invades underlying tissue sometimes including bone, and which rarely metastasizes to the regional lymph nodes. Histological examination shows a well-differentiated squamous epithelial proliferation without cellular atypia, with numerous crypts and sinuses. The diagnosis is difficult as the cytological regularity does not allow malignancy to be evoked. It is the conspicuity of the invasion of the deep tissues with sufficiently large samples that makes it certain. The differential diagnosis includes keratoacanthoma, pseudo-epitheliomatous hyperplasia mainly represented by pyoderma vegetans and "classical" squamous cell carcinoma. From a nosological point of view the authors totally assimilate Epithelioma cuniculatum and Gottron's papillomatosis cutis carcinoïdes regrouped under the name of cutaneous verrucous carcinoma, emphasizing the analogy with verrucous carcinomas of mucous membrane, giant condyloma of Buschke-Lowenstein, oral florid papillomatosis.

[A case of malignant melanoma expressing epithelial cell markers in immunohistochemistry using paraffin sections]. / Un cas de mélanome malin exprimant les marqueurs des cellules épithéliales en immunohistochimie sur coupes en paraffine.

Recently, applications of immunohistochemical techniques for the cytoplasmic localization of intermediate filaments has produced advances in tumor diagnosis and characterization. We report a 58-year-old white male with a clinically and histologically typical metastatic malignant melanoma. This case was peculiar because the same neoplastic cells stained for both S-100 protein and keratin on paraffin embedded tissue. These facts illustrate how cautiously we must interpret the positivity of immunohistochemical technics. We thus insist on the importance of the clinical and basic histologic data for the diagnosis, in order to avoid errors.

[The association of neuro-endocrine carcinoma of the skin and Bowen's disease. Review of the literature apropos of 4 cases]. / Association d'un carcinome neuro-endocrine cutané et d'une maladie de Bowen. Revue de la littérature à propos de quatre cas.

Four cases of neuroendocrine carcinoma following Bowen's disease are presented. An immunohistochemical study was performed. The four patients, 3 men and a woman, ranging from seventy to eighty-seven years of age, developed a nodular tumor on a preexisting cutaneous lesion. In one of those cases the diagnosis of Bowen's disease was confirmed histologically before the apparition of the nodular tumor. The tumors were localized on the scalp, thorax, dorsum of the hand, and the scrotum. The four tumors were immunohistologically typical of neuro-endocrine carcinoma: there was a positivity for neurofilaments, cytokeratins and neurone-specific enolase. The clinico-pathological characteristics of those 4 neuro-endocrine carcinomas associated with a Bowen's disease, when compared with the 15 similar described in the literature, are identical to the isolated neuroendocrine carcinoma, from a clinical, morphological and evolutional point of view. The majority are seen in patients older than 60 years old and one third of the cases described survived at least 5 years. The coexistence of Bowen's disease and neuroendocrine carcinoma, the association of neuroendocrine and epidermoid cells in other cutaneous tumors, reactivate the controversy concerning the histogenesis of the so-called Merkel cell carcinoma. In fact, the histogenesis of the tumor is still not fully understood.

[Lyme disease]. / La maladie de Lyme.

The history of Lyme disease, a contagious condition caused by Borrelia burgdorferi transmitted to man by ticks offers infectiologists a formidable lesson on how medicine progresses. Clinical description started in Europe at the turn of the century with Pick's description of what was then labelled chronic atrophic acrodermatitis. Fifty years later Hauser noted the affection was transmitted by ticks. Independently, Afzelius, then Lipschutz, described erythema chronicum migrans and its relationship with tick bites. Neurological involvement was also described with the skin signs. These early dermatological descriptions suddenly came into the limelight in 1975 when an epidemia of arthritis occurred in children in Lyme, Connecticut, USA. Many of the affected children had erythema chronicum migrans. Based on these observations and an epidemiological analysis of the epidemia, Steele and co-workers defined "Lyme disease" as a rheumatological disorder commonly associated with erythema chronicum migrans and sometimes with multiple organ involvement. In 1982 Borgdorfer suggested that tick bites transmitted a Spirochaeta which was later authentified as the causal agent: Borrelia burgdorferi. Immunofluorescence and ELISA tests were rapidly developed for the diagnosis of infection by this germ which is very difficult to culture. Antibiotic curative treatment was immediately available and in 1991 a consensus conference established recommendations for treatment of isolated and disseminated forms. Antibiotic prophylaxis is not necessary but rapid extraction of the tick after the bite can prevent the disease as transmission from tick to man takes several hours. And medical progress continues. Work is now being conducted on evaluating the extent of late neurological manifestations, on developing polymerase chain reaction methods to identify B. burgdorferi infection in specific organs and on developing a vaccine.

[Paraneoplastic pemphigus]. / Pemphigus paranéoplasique.

A RARE DISEASE: Paraneoplastic pemphigus is an rare autoimmune bullous skin disease recently recognized. About 50 cases have been reported since its first description in 1990. CLINICAL MANIFESTATIONS: Clinical signs are polymorphous resembling the cutaneomucosal manifestations of pemphigus vulgar (skin and mucosa erosions, fragile interdermal bullae), pemphigoid (urticaria, distended subepidermal bullae), and polymorphous erythema (plaque lesions). Mucosal erosions predominate however. ASSOCIATED CANCERS: Most cancers associated with paraneoplastic pemphigus are hematologic diseases (non-Hodgkin's lymphomas, chronic lymphoid leukemia). SEVERE PROGNOSIS: No standard treatment has been defined. General corticosteroids and treatment of the causal disease are indicated. The clinical course of paraneoplastic pemphigus does not always follow the course of the associated neoplasm. POSITIVE DIAGNOSIS: Pathology criteria (keratinocyte necrosis, suprabasal keratinocyte vacuolization, intraepidermal acantholysis) and immunohistological findings (antibody and complement deposits at the dermo-epidermal junction and within the keratinocytes on different epithelial substrates) are insufficient for positive diagnosis. Autoantibodies must be identified by immunoprecipitation or immunoblotting to identify the target antigen complex plakin components (desmoplakin I and II, periplakin, envoplakin), the major pemphigoid antigen, desmoglein 3, and certain yet unidentified antigens with a molecular weight of 170 kD. PATHOGENESIS: Paraneoplastic pemphigus appears as a model autoimmune paraneoplastic disease. Its origin remains elusive. It has been hypothesized that tumor-induced inhibition of tolerance to certain antigens implicated in the keratinocyte junctional systems could be involved.

[Bacillary angiomatosis]. / Angiomatose bacillaire.

Bacillary angiomatosis (BA) is a recently described infection usually found in patients with human immunodeficiency virus disease. BA is caused by a Gram-negative coccobacillus. This organism is primarily responsible for skin lesions of the pseudo-botryomycoma type or inflammatory nodules, but it also produces fever, degradation of the general condition and visceral lesions involving the lymph nodes, the liver, the spleen and the bones. Histology shows vascular proliferation with turgid endothelial cells and mostly neutrophilic inflammatory infiltrates. BA is susceptible to many antibiotics. The authors describe the history of the disease and its clinical and histological features, discuss its differential diagnosis and principally deal with the relationship between BA and cat-scratch disease and between BA and verruca peruana. They also present the molecular biology technique which enables a genotypic diagnosis of the disease to be made, replacing a deficient phenotype.

[Pyoderma gangrenosum]. / Le pyoderma gangrenosum.

GENERAL CHARACTERISTICS: Rare cause of cutaneous ulceration, pyoderma gangrenosum is among the group of neutrophilic dermatites characterized by the richness of the mature neutrophilic polynuclear dermal infiltrate. The primary lesion is a pustule sitting on an inflammatory base; extremely painful, it rapidly ulcerates. The lesion rapidly extends to more than 10 cm in diameter, has a regular, sharp border and a peripheral roll of flesh exhibiting purulent channels on the inside and a red granulous surface often covered with a pustular coating. Little is known of this disease. CONCOMITANT AFFECTIONS: In more than 50% of cases, pyoderma gangrenosum is associated with other diseases, which must be systematically searched for. These may be digestive, essentially inflammatory enterocolitis with frequent development of peristomal ulceration, rheumatismal affections notably rheumatoid arthritis, hematological affections (benign monoclonal gammapathy, chronic myeloid hemopathy). FROM A PARACLINICAL POINT OF VIEW: There are no specific examinations. A cutaneous biopsy should be performed in all cases, notably to eliminate other causes of ulceration. Since concomitant disease can be subsequently revealed, it is essential to renew the paraclinical investigations, even after the disease has healed. NO CODIFIED TREATMENT: Treatment of the cause, if it can be cured, may be sufficient to permit regression of the lesions. Local treatments to provoke budding and hence avoid surinfection are mandatory. In the progressive and extensive forms, systemic treatment, notably high dose corticosteroids, is indicated. Surgery, a priori, is excluded.

[Chronic lupus erythematosus in lupus disease. Retrospective study of 136 patients]. / Place du lupus érythémateux chronique au sein de la maladie lupique. Etude rétrospective de 136 patients.

OBJECTIVES: The aim of this retrospective study of 136 patients was to specify the natural history and the systemic prognosis of chronic cutaneous lupus erythematosus. It has been stated that in most of the cases, the disease only affects the skin. METHODS: From 10 October 1980 to 31 December 1990, 136 patients with the following criteria were included in this retrospective study: clinical signs suggestive of chronic cutaneous lupus erythematosus, characteristic histology, insufficient evidence for the diagnosis of systemic lupus erythematosus. RESULTS: The prevalence of systemic clinical involvement in the population under study was nearly the same as in the general population. The following biologic or immunologic abnormalities were quite common: leukopenia, lymphopenia, thrombopenia (significantly more frequent among patients having widespread chronic cutaneous lupus erythematosus), low titers of complement levels, positive antinuclear antibodies (usually at a low titer). Eleven out of the 136 patients developed systemic lupus erythematosus, most often over 5 years after the onset of the cutaneous lesions. Four cases out of these 11 had poor prognosis: renal and/or neurologic involvement. CONCLUSION: This data suggests that patients with chronic cutaneous lupus erythematosus could benefit from long-term follow up, since the course to systemic disease occurs in only a few. Usually, antimalarials used singly or in combination with topical steroids may lead to the clearing of the lesions. Thalidomide will occasionally be useful whenever the disease is unresponsive to the preceding measures.

[Cowden's disease or the multiple hamartoma syndrome]. / La maladie de Cowden ou syndrome des hamartomes multiples.

Cowden's disease, also called multiple hamartoma syndrome, is a clinical entity characterized by hamartomatous tumours of endodermal, mesodermal and ectodermal origin. Although extremely rare, the disease must be known to all internists. A case of Cowden's disease in a 36-year old male patient is reported. The authors insist on the high incidence of digestive disorders and the risk of malignant degeneration of mammary and thyroid tumours. They also describe the cutaneous and mucosal lesions characteristic of the disease.

[Idiopathic eruptive macular pigmentation (author's transl)]. / La pigmentation maculeuse éruptive idiopathique.

The feeling of the authors is that their seven reported cases of a pigmented dermatosis are different from the ashy dermatosis and from the erythema dyschromicum perstans. This disease, which affects children and teenagers, males as well as females, is characterized by pigmented macules 5-25 mm in diameter, affecting the neck, the trunk and the limbs. The first symptom is whether a pigmented spot, or an erythematous, papular or achromic lesion; in the latter instance the pigmentation occurs only secundarily. In most of the cases this dermatose is slowly and spontaneously regressive. The histological picture is not really specific. In one case there was a marked intraepidermal dyskeratosis of the sweat duct openings. The etiology remains unknown.

[Follicular tumidus retro-auricular lichen planus (author's transl)]. / Lichen plan folliculaire tumidus rétro-auriculaire

Referring to three comparable cases, a new form of lichen planus is described. This form is characterized by a prominent lesion, violaceus in color or pigmented, with white yellowish specks mimicking milia. The histologic picture of lichen follicularis, the presence of typical papules of lichen distant from the retro-auricular lesion make it possible to consider this clinical aspect as a variety of lichen planus.