Tofacitinib for celiac disease and microscopic colitis: killing two birds with one stone.

Microscopic colitis is a chronic inflammatory condition of the colon. Firstline treatment consists of budesonide, with the consideration of biological agents in refractory cases. Celiac disease is a chronic immune mediated and gluten-induced enteropathy, with treatment consisting of a gluten-free diet. There is an association between microscopic colitis and instead of xand celiac disease, especially in refractory cases they can coincide. In this manuscript, we report for the first time the efficacy of tofacitinib, a pan Janus kinase inhibitor, in the treatment of concomitant microscopic colitis and celiac disease, resulting in persistent clinical and histological remission.

[Idiopathic intracranial hypertension: From physiopathological mechanisms to therapeutic decision]. / Hypertension intracrânienne idiopathique : des mécanismes physiopathologiques à la décision thérapeutique.

Clinical features include visual disturbances, headaches, and pulsatile tinnitus that can be associated with reduced quality of life, and a risk of irreversible visual impairment in some cases. Obese women of childbearing age represent the main at-risk population, and the incidence of the disease is increasing because of rising prevalence of obesity worldwide. In addition, an imbalance in sex hormones is reported as a contributing risk factor. The pathophysiology of idiopathic intracranial hypertension involves a disturbance of the evacuation pathway of intracranial fluids caused by the increase in intracranial venous pressure. Brain imaging is useful for diagnosis with several signs including bilateral stenosis of the transverse sinuses that plays a major role in the pathogenesis of the disease by creating a positive feedback loop that increases intracranial venous hypertension and contributes to clinical manifestations. Treatment aims to relieve symptoms and prevent permanent visual impairment. Drug therapies including acetazolamide and topiramate have moderate effectiveness. Among invasive treatments, transverse sinus stenting seems to be the most interesting option to consider in drug-resistant patients. Weight loss remains essential to achieve a sustainable improvement by reducing central venous pressure. Future randomized trials are expected to reach a consensus on this treatment.

Whipple's disease in a man of North African descent : case report and brief review of the literature.

A 62-year-old man of North African descent presented with weight loss in the past year and diarrhea for three weeks. His medical history included erosive rheumatoid arthritis, treated with methotrexate and adalimumab. Histological examination of a duodenal biopsy showed foamy macrophages in the lamina propria, with PAS-positive cytoplasmatic inclusions. These findings are compatible with Whipple's disease, a rare chronic infectious disease caused by Tropheryma whipplei, an opportunistic bacterium. It is typically seen in middle-aged Caucasian men and the immunocompromised host. The classical presentation of Whipple's disease consists of intermittent migratory arthralgia, followed by intestinal symptoms which typically occur six to seven years later. The clinical image can be very variable, and this complicates the diagnostic process. PAS-staining and PCR are the diagnostic cornerstones. In our case, treatment consisted of a prolonged cure of antibiotics: intravenous ceftriaxone for two weeks, followed by an oral maintenance therapy of doxycycline and hydroxychloroquine for at least one year. A therapeutic dilemma arose as continued anti-TNF blockade was necessary to maintain remission of the rheumatoid arthritis. Lifelong follow-up is necessary because relapse is possible.

Acute angiotensin-converting enzyme inhibition increases the plasma level of the natural stem cell regulator N-acetyl-seryl-aspartyl-lysyl-proline.

Angiotensin I-converting enzyme (ACE) has two homologous active NH2- and COOH-terminal domains and displays activity toward a broad range of substrates. The tetrapeptide N-acetyl-seryl-aspartyl-lysyl-proline (Ac-SDKP) has been shown to be hydrolyzed in vitro by ACE and to be a preferential substrate for its NH2-terminal active site. This peptide is a regulatory factor of hematopoiesis which reversibly stem cells and normal early progenitors into S-phase. We found that a single oral dose of 50 mg of the ACE inhibitor, captopril, when administered to eight healthy subjects in a double-blind, crossover, placebo-controlled study, massively increased the plasma level of Ac-SDKP. ACE inhibition by captopril induced a 90-99% inhibition of in vitro [3H]Ac-SDKP hydrolysis and a long-lasting 5.5-fold (range: 4-8.5-fold) increase in the plasma levels of Ac-SDKP. These results demonstrate that Ac-SDKP is the first natural peptide hydrolyzed by the NH2-terminal domain of ACE not only in vitro but also in vivo, confirming that both catalytic sites of ACE are physiologically active. Our data suggest that ACE may also be implicated in the process of hematopoietic stem cell regulation, by permanently degrading this natural circulating inhibitor of cell entry into S-phase.

Nitroglycerin metabolism by Phanerochaete chrysosporium: evidence for nitric oxide and nitrite formation.

We have demonstrated that a filamentous fungus Phanerochaete chrysosporium converts glyceryl trinitrate (GTN) into its di- and mononitrate derivatives concurrently with the formation of nitric oxide detected by electron paramagnetic resonance (EPR), and the formation of nitrite. The metabolisms of nitrite and nitrate by the fungus are evaluated and taken into account when considering GTN degradation. Lack of evidence for nitrate formation from GTN suggests that an esterase-type activity is not involved. Furthermore, the kinetics of appearance of the hemoprotein-NO and non-heme protein-NO (FeS-NO) complexes indicate that an enzymatic process producing NO directly from GTN may be involved concurrently with a glutathione transferase-like system.

Further purification of bovine spleen inhibitors of lymphocyte DNA synthesis (chalones).

Partially purified lymphocytic factors were obtained from boving spleen; these factors are non-cytotoxic and biologically active in vitro and in vivo: [3H]Thymidine incorporation into DNA of mitogen-stimulated mouse spleen cells in culture is inhibited; similar results are obtained with phytohaemagglutinin-stimulated peripheral human lymphocytes, where blast cells transformation is blocked. [3H]Thymidine incorporation into DNA of mitogen stimulated lymphocytes withdrawn from in vivo treated mice, is also reduced. The two factors in vitro and in vivo seem to act preferentially on mouse spleen cells compared to their action on liver, kidney and testicle cells in cluture, as far as thymidine incorporation into DNA is concerned. The following techniques were applied for their purification: 1. Homogenization of the fresh tissue in water, centrifugation, dialysis of the supernatant, centrifugation, fractionation of the supernatant by alcoholic precipitation and finally concentration in vacuo and lyophilization of the material soluble at 75% of alcohol yielded fraction F. 2. Preparation of an acetone powder from the spleen, extraction of the dry powder with water, then high speed centrifugation, followed by lyophilization of the supernatant produced fraction F'. Both fractions F and F' were further fractionated by chromatography on a Sephadex G75 column: 7 peaks were obtained (F1--F7). Biological activity was found in fraction F1, corresponding to high molecular weight material, and in fraction F6, corresponding to low molecular weight substances. By rechromatography on Sephadex G 75, it is easy to dissociate from F1 a small molecular weight fraction which might be similar to F6 as far as elution volume and biological properties are concerned.

A new approach of urodele amphibian limb regeneration: study of myosin isoforms and their control by thyroid hormone.

In P. waltlii, an urodele amphibian species which undergoes spontaneous metamorphosis, study of native myosin in pyrophosphate gels at various stages of normal development demonstrates a complete larval to fast myosin isoforms transition, which occurs more precociously in forelimb muscles than in the dorsal and ventral muscles. In the neotenic species A. mexicanum, forelimb muscles development also presents a complete myosin isoforms transition which is in contrast with the partial myosin isoforms transition observed in the dorsal muscle. In metamorphosed or neotenic animals of both species aged 1 year, forelimb regeneration is characterized by a complete transition from larval to fast myosin isoforms, that occurs earlier and more rapidly than in normal forelimb development. When forelimb regeneration is studied in P. waltlii aged 4 years, the adult fast and slow isomyosins are expressed very early in the regeneration process. In experimental hypothyroidian P. waltlii, the larval to fast isoforms transition in regenerating forelimb muscles is slightly delayed. Experimental hyperthyroidism accelerates the disappearance of larval isomyosins in regenerating forelimb muscles, both in P. waltlii and A. mexicanum aged 1 year. This work demonstrates that changes in myosin isoform pattern during forelimb regeneration in adult urodele amphibians are different from changes occurring in the normal forelimb development. They take place without any thyroid hormone influence, as opposed to normal development, and appear to be age-dependent.

AcSDKP, an inhibitor of CFU-S proliferation, is synthesized in mice under steady-state conditions and secreted by bone marrow in long-term culture.

The peptide AcSDKP, isolated from fetal calf bone marrow, is able to prevent DNA synthesis in mouse CFU-S in vivo and in vitro. The molecule is demonstrated here to be constitutively produced in mice and synthesized by bone marrow cells in long term culture.

Lisinopril, an angiotensin I-converting enzyme inhibitor, prevents entry of murine hematopoietic stem cells into the cell cycle after irradiation in vivo.

The hemoregulatory peptide N-Acetyl-Ser-Asp-Lys-Pro (AcSDKP) has been shown in vivo to inhibit the cycling of murine hematopoietic stem cells triggered into S-phase by either cytotoxic drug administration or irradiation. This property, further confirmed using in vitro models, demonstrates that the peptide has an in vivo protective effect on the hematopoietic system. AcSDKP has been shown to be a physiological substrate of angiotensin I-converting enzyme (ACE), which catabolizes the peptide through a dipeptidasic activity. Thus, oral administration of ACE inhibitor to humans has led to an increase in the plasma AcSDKP concentration. In the present paper, we report on the in vivo effect of lisinopril, an ACE inhibitor, on the proliferative status of murine hematopoietic stem cells triggered into S-phase by irradiation. Administration of lisinopril (10 mg/kg) 1 hour after irradiation led to a 90 to 100% inhibition of murine plasma ACE activity as observed during the first 4 hours postirradiation. This inhibition was correlated with a 600% increase in the endogenous plasma AcSDKP level and a total suppression at 24 hours of entry of the hematopoietic stem cell into the cell cycle. We discuss the possible role of ACE in the regulation of hematopoietic stem cell proliferation through control of the AcSDKP concentration.

Enhancement of the adherence of hematopoietic stem cells to mouse bone marrow-derived stromal cell line MS-1-T by a tetrapeptide acetyl-N-Ser-Asp-Lys-Pro.

The tetrapeptide acetyl-N-Ser-Asp-Lys-Pro (AcSDKP) has been shown to inhibit in vivo the hematopoietic stem cell (spleen colony-forming unit; CFU-S) entry into cell cycle in cytosine arabinoside-treated mice. Our data showed that AcSDKP has no effect on interleukin 3 (IL-3)-dependent DA-1 cell proliferation or on granulocyte-macrophage colony-forming cell and mast cell colony formation, whereas it enhances the adherence of CFU-S to the bone marrow-derived hematopoietic supportive stromal cell line MS-1-T. AcSDKP suppresses MS-1-T proliferation but does not modify granulocyte-macrophage colony-stimulating activity secretion by these cells. This suggests that the peptide does not counteract the activity of the IL-3 receptor on CFU-S but acts on MS-1-T and in particular at the level of CFU-S/MS-1-T interactions.

Pituitary-thyroid axis controls the final differentiation of the dorsal skeletal muscle in urodelan amphibians.

A histoenzymological study of the ATPase activity of myosin in the dorsal axis muscle (dorsalis trunci) was carried out on two species of urodelan amphibians: Pleurodeles waltlii, a euthyroid species with spontaneous metamorphosis and Ambystoma mexicanum, a neotenic hypothyroid species. P. waltlii and A. mexicanum underwent an operation after which cytological analysis of the remaining pituitary were carried out in parallel. The muscle phenotype of urodelan amphibians varies according to the thyroid status of the species. In euthyroid adults, IIA fibers are dominant whereas in hypothyroid adults, IIC fibers are dominant. The number of type IIB (fast) and type I fibers (slow) are similar in both species. Physiological or experimental modulation of the concentration of circulating thyroid hormones results in a modification of the muscle fiber type profile pertaining to the considered species. We found that pituitary (TSH) plays a dominant role in the maturation of type IIC fibers in both species. Moreover, it seems to modulate the development of IIA fibers in P. waltlii and that of IIB fibers in A. mexicanum. Its action is thus species specific. Through partial or total hypophysectomy experiments, we have been able to demonstrate the influence of the hypophysothyroidian axis on the appearance of the adult muscle phenotype during metamorphosis.

The emergence of an adult muscle phenotype in urodelan amphibians: an immunohistochemical study.

Electrophoretic techniques adapted for the analysis of muscles of lower invertebrates reveal four myosin heavy chain isoforms in the dorsalis trunci of Pleurodeles waltlii: two fast (MHC-IIA, MHC-IIB), and one slow (MHC-I) in the adult and one isoform (MHC-La) in the larvae. Polyclonal antibodies were prepared against the larval (anti-MHC-La) and one of the fast myosin (MHC-IIA) isoforms and their specificity was confirmed by western blot analysis. An immunohistochemical analysis was then carried out on frozen sections of the dorsalis trunci of P. waltlii at different stages of development. From stage 44 it was possible to demonstrate the presence of MHC-IIA in the small diameter fibers at the periphery of the muscle; the number and diameter of these fibers increased from stage 44 to stage 56 when anatomical metamorphosis had finished. By stage 56 these fibers could also be readily identified using standard histochemical techniques as type IIA fibers. We conclude that fast IIA myosin is expressed well before the final adult muscle phenotype has been established and its expression is therefore independent of thyroid hormone.

Myosin structure and thyroidian control of myosin synthesis in urodelan amphibian skeletal muscle.

Electrophoretic analysis in non-dissociating conditions reveals three types of myosin in adult urodelan amphibian skeletal muscles: 3 isoforms of fast myosin (FM), one isoform of intermediate myosin (IM) and one or two isoforms of slow myosin (SM). Each type is characterized by a specific heavy chain HCf (FM), HCi (IM) and HCs (SM), respectively. In all urodelan species, as in mammals, fast isomyosins associate HCf and the three fast light chains LC1f, LC2f, and LC3f. In most urodelan species the intermediate myosin contains LC1f and LC2f and can be considered as an homodimer of the alkali LC1f. However, in Euproctus asper, IM is characterized by the association of both slow and fast LC with HCi. Slow myosin is a hybrid molecule associating HCs with slow and fast LC. During metamorphosis, a myosin isoenzymic transition occurs consisting in the replacement of three larval myosins (LM) characterized by a specific heavy chain (HCI), by the adult isomyosins with lower electrophoretic mobilities. At the same time there is a change in the ATPase myofibrillar pattern, with the larval fiber types being replaced by adult fibers of types I, IIA and IIB. In the neotenic and perennibranchiate species, which do not undergo spontaneous metamorphosis, sexually mature larval animals present a change in the myosin isoenzymic profile, but no complete transition. The coexistence of larval and adult isomyosins and the persistence of transitional fibers of type IIC in the skeletal muscle are demonstrated. Experimental hypo- and hyperthyroidism indicate that thyroid hormone stimulates the regression of the larval isomyosins, possibly through indirect pathways. In contrast, the appearance and the persistence of the adult isomyosins seem to be independent of thyroid hormone. Thus, the control of the isoenzymic transition in the skeletal muscle of urodelan amphibians appears to imply indirect mechanisms, operating differently on each of the two phases of the complete transition.

The mechanism of action of the tetrapeptide acetyl-N-Ser-Asp-Lys-Pro (AcSDKP) in the control of haematopoietic stem cell proliferation.

The mechanism of action of the haemoregulatory tetrapeptide Acetyl-N-Ser-Asp-Lys-Pro (AcSDKP, M(r) = 487 amu), was investigated using an in vitro assay of a murine high proliferative potential colony-forming cell (HPP-CFC) which responds to proliferation regulators of the haematopoietic stem cell population. AcSDKP had no direct inhibitory effect on the number, or the proportion in S phase, of the committed granulocyte-macrophage progenitor (GM-CFC), or cycling HPP-CFC populations. However, AcSDKP blocked the action of a stimulator of haematopoietic stem cell proliferation, preventing the switching of quiescent HPP-CFC into cell cycle. It would appear that AcSDKP exerts its inhibitory haemoregulatory role indirectly, by preventing stimulator action on haematopoietic stem cells.

Involvement of thymosin beta 4 and endoproteinase Asp-N in the biosynthesis of the tetrapeptide AcSerAspLysPro a regulator of the hematopoietic system.

It is shown that AcSDKP a new regulator of the hematopoietic system can be generated from thymosin beta 4 by a one-step enzymatic cleavage in vitro and in vivo. AcSDKP and T beta 4 were both detected in bone marrow cells (BMC). Incubation of [3H]T beta 4 with either intact or lysed BMC led to the formation of [3H]AcSDKP whereas the labelled tetrapeptide was not degraded under these conditions. Model enzymatic degradation of T beta 4 carried out with bacterial enzymes suggests that a mammalian endoproteinase Asp-N might be involved in the formation of AcSDKP through the specific cleavage of the 4Pro-5 Asp peptide bond of T beta 4.

Myosin structure in the eel (Anguilla anguilla L.). Demonstration of three heavy chains in adult lateral muscle.

Myosin extracts from central white fibers and peripheral red fibers of the lateral muscle of eel (Anguilla anguilla) were analysed by electrophoresis under non-dissociating conditions, which demonstrated a polymorphism of myosin isoforms. The light and heavy subunit content of the isomyosins was established using SDS-PAGE and two-dimensional electrophoresis. In the central white muscle, 3 myosin isoforms FM3, FM2, FM1, were characterized by 3 types of fast light chain and one fast heavy chain HCf; the existence of a fourth isomyosin is discussed. In the peripheral red muscle, two myosin isoforms were found, SM1 and SM2, each characterized by a specific heavy chain, HCs1 or HCs2, and containing the same slow light chain content. This work demonstrates for the first time the existence of 3 heavy chains in the skeletal muscle of a fish.

[Incorporation of tritiated thymidine into DNA of rat liver: a critique of various evaluation methods]. / Incorporation de la thymidine tritiée dans le DNA de foie de rat: critique des différentes méthodes d'évaluation

We have measured the incorporation of 3H-(methyl)-thymidine by cell cultures of rat foetal liver and in vivo by the livers of young rats stimulated by casein, in order to compare three methods for the extraction of DNA. The DNA was extracted by three different techniques: perchloric acid precipitation, trichloroacetic acid precipitation and phenol extraction, and its specific activity was determined. The radioactive labelling was also determined for the lipid, ribonucleic acid and protein fractions for the two first methods, in both of which 70 p. cent of the incorporated tritium is found in the DNA fraction and about 10 p. cent in each of the other fractions. The determination of the specific radioactivity of DNA gives similar results for the three extraction methods. However, since larger yields were obtained by both acid precipitation techniques than by phenol extraction, we believe them to be more suitable for studies on cell cultures.

Whole abdominal irradiation following chemotherapy in patients with minimal residual disease after second look surgery in ovarian carcinoma.

From January 1981 through December 1985, 65 patients with epithelial carcinoma of the ovary were treated with the following protocol: surgery, combination chemotherapy, second-look surgery documenting tumor less than or equal to 2 cm, and whole abdominal irradiation. Chemotherapy consisted of a combination of cyclophosphamide, adriamycin, and cisplatinum in 89% of the patients. The median number of cycles was eleven. Second-look surgery documented no residual tumor in 23 patients, microscopic disease in three patients, and macroscopic disease less than or equal to 2 cm in 39 patients. Whole abdominal irradiation was given with an open field technique up to 20 Gy without renal or hepatic shield. A pelvic boost of 15-30 Gy was subsequently added in 17 patients with macroscopic disease in the pelvis at the time of second-look surgery. Fifteen patients received complementary chemotherapy mostly hexamethylmelamine. All but two patients completed whole abdominal irradiation: one refused further radiotherapy after 3 Gy and one developed disease progression with bowel obstruction after 1 Gy. The median follow-up was 69 months. The 3-year and 6-year no evidence of disease survival rates were 60% (95% CI: 48-71) and 33% (95% CI: 21-46), respectively. The 3-year and 6-year recurrence rates were 33% (95% CI: 22-45) and 54% (95% CI: 40-67), respectively. The 3-year and 6-year metastasis rates were 22% (95% CI: 13-34) and 43% (95% CI: 30-58), respectively. A multivariate analysis showed that residual disease after second-look surgery was the only significant prognostic factor with a relative risk of death or local or distant failure of 4.2 (95% CI: 1.9-9.5, p less than 10(-4)). Two patients developed mean-term gastrointestinal complications (small bowel obstructions requiring surgery). Survival remains poor with high level of failure even with aggressive multimodal treatment.

Prognostic value of hemoglobin concentrations and blood transfusions in advanced carcinoma of the cervix treated by radiation therapy: results of a retrospective study of 386 patients.

A retrospective study was carried out on 386 patients with advanced cervical carcinomas treated with radiation therapy between 1973 and 1983. The influence of hemoglobin concentrations and blood transfusions before and/or during treatment on the occurrence of distant and/or local regional failures were examined in a univariate and multivariate analyses. In the multivariate analysis hemoglobin concentrations were prognostic only during treatment and patients with at least one value below the threshold of 10 gm% had a significantly higher risk of local regional failure than the patients with all their values above the threshold. Moreover 70% of these high risk patients had less than half of their values below the threshold. It is possible that blood transfusions might be beneficial when given before treatment. However, although it was not significant, blood transfusions given during treatment tended to be an adverse prognostic factor suggesting that blood transfusions might not have completely offset acute anemia prior to transfusion. Our study suggests that anemia during treatment, even of short duration might be detrimental to patients.

Synthesis and activity of NAcSerAspLysPro analogues on cellular interactions between T-cell and erythrocytes in rosette formation.

Analogues of NAcSerAspLysPro (AcSDKP), a natural regulator of hematopoiesis isolated from fetal calf bone marrow, were synthesized. The biological activity of these molecules were evaluated in vitro in the rosette assay, which measures the interaction between human Jurkat T-cells and sheep red blood cells. In this test, the tripeptide SerAspLys was the most efficient. Inhibitory activity was detected at the concentration 10(-14) M for the analogue and at 10(-9) M for the parent tetrapeptide. The dipeptide NAcSerAsp still showed activity but at much higher doses (10(-6) M). Substitution of polar amino acids led mostly to inactive molecules. Thus, replacement of Ser by Ala, or Lys by Orn yielded completely inactive compounds and replacement of Asp by Glu decreased the activity (10(-6) M). The present study gives an insight into the role of individual amino acids of AcSDKP in the inhibition of the rosette formation which implicates interactions with T-cell CD2 glycoprotein.