Cementum protein 1 transfection does not lead to ultrastructural changes in nucleolar organization of human gingival fibroblasts.

BACKGROUND AND OBJECTIVE: Transfection of cementum protein 1 (CEMP1) into human gingival fibroblasts (HGFs) notably increases cell metabolism and results in overexpression of molecules related to biomineralization at transcriptional and protein levels. Therefore, HGF-CEMP1 cells are considered as putative cementoblasts. This represents a significant advance in periodontal research because cementum neoformation is a key event in periodontal regeneration. In addition, it is well known that important changes in cell metabolism and protein expression are related to nucleolar structure and the function of this organelle, which is implicated in ribosome biogenesis. The aim of this study was to determine the effect of transfecting CEMP1 gene in human HGF on the ultrastructure of the nucleolus. MATERIAL AND METHODS: Cells were processed using the conventional technique for transmission electron microscopy, fixed with glutaraldehyde, postfixed with osmium tetraoxide, and embedded in epoxy resin. Semi-thin sections were stained with Toluidine blue and observed by light microscopy. Thin sections were stained with uranyl acetate and lead citrate. For ribonucleoprotein detection, the staining method based on the regressive effect of EDTA was used. In addition, the osmium ammine technique was used for specific staining of DNA. RESULTS: The results obtained in this study suggest that transfection of CEMP1 into HGFs does not produce changes in the general nucleolar ultrastructure because the different components of the organelle are present as fibrillary centers, and dense fibrillar and granular components compared with the control. CONCLUSION: The transfection of CEMP1 into HGFs allows these cells to perform cementoblast-like functions without alteration of the ultrastructure of the nucleolus, evaluated by the presence of the different compartments of this organelle involved in ribosomal biogenesis.

Usage and attitudes of physicians in Japan concerning traditional Japanese medicine (kampo medicine): a descriptive evaluation of a representative questionnaire-based survey.

Kampo medicine has been the primary medical model in Japan until the mid 1800s, regained a prominent role in today's Japanese medical system. Today, 148 herbal Kampo formulas can be prescribed under the national health insurance system, allowing physicians to integrate Kampo in their daily practice. This article aims to provide information about the extent to which Kampo is now used in clinics throughout Japan and about physician's current attitudes toward Kampo. We used the results of a 2008 survey that was administered to physicians throughout Japan (n = 684). The data showed that 83.5% of physicians currently use Kampo in the clinic, although the distribution of physicians who use Kampo differ widely depending on the specialty and provided a breakdown of Kampo usage by specialty. It will be interesting to see how each specialty incorporates Kampo into its respective field as Kampo continues to play a pertinent role in Japanese medical system.

Autoimmune associations in a Mexican cohort with primary biliary cholangitis. / Asociaciones autoinmunes en una cohorte mexicana con colangitis biliar primaria.

INTRODUCTION: Several groups have reported associations of primary biliary cholangitis with other autoimmune entities, particularly Sjögren's syndrome and hypothyroidism. Its prevalence and characteristics in Mexican patients is unknown. AIM: To determine the frequency and characteristics of autoimmune diseases in a Mexican cohort of patients with primary biliary cholangitis. MATERIALS AND METHODS: The medical records of patients that presented with primary biliary cholangitis within the time frame of 2005 and 2012 were reviewed and assessed for other autoimmune diseases. RESULTS: Seventy-eight patients, 75 women and 3 men, were included. Their mean age was 55.8 years. Seventy-three cases had positive antimitochondrial antibodies (94.8%) and disease was confirmed in 5 through liver biopsy. Five patients (8%) had anti-smooth muscle antibodies and 55/78 (70.5%) had antinuclear antibodies by indirect immunofluorescence. Forty-nine patients (62.8%) were positive for an autoimmune disease other than primary biliary cholangitis. Among those, 20 patients had one associated disease, 14 had 2, and 15 patients had 3 concomitant diseases. They included: Sjögren's syndrome in 23/78 patients (29.5%), dysthyroidism in 21/78 cases (26.9%), Raynaud syndrome in 11/78 (14.1%), CREST syndrome in 9/78 patients (11.4%), rheumatoid arthritis in 6/78 patients (7.7%), vitiligo in 5/78 (6.4%), scleroderma in 4/78 patients (5.1%), and other diseases in 8 patients. In 12/78 patients (15.4%), there was a documented family background of autoimmune disease. CONCLUSIONS: The presence of autoimmune associations in our cohort was frequent, and similar in characteristics to the information reported by other groups. The clinical implications of those findings remain to be determined.

Early and late fistula failure.

While an arteriovenous fistula is the best available form of hemodialysis access, a significant number of fistulae never mature to support dialysis (early failure) or fail after successful use (late failure). Venous stenosis and the presence of accessory veins are the two main causes of early failure. Recent data have demonstrated that a great majority of such AVFs can be successfully salvaged by percutaneous interventions and become available for dialysis. In addition to early failure, a great majority of thrombosed fistulae can also be successfully declotted using simple endovascular techniques. Fistula thrombosis has clear differences from graft clotting. First of all, cannulation of a clotted fistula is more challenging. Secondly, thrombus volume present in a clotted fistula can be quite variable. A fistula might thrombose with minimal or no thrombus. At other times, there is moderate-to-severe thrombus burden that accompanies fistula clotting. While percutaneous balloon angioplasty to correct the underlying stenosis might be all that is needed to declot a fistula with no thrombus, thromboaspiration is required to successfully declot a fistula with moderate thrombus. Salvage of early and late fistula failure is critical to minimize catheter use and is supported by the National Kidney Foundation Dialysis Outcomes Quality Initiative. Additionally, it is a powerful strategy to maximize AVF use in hemodialysis patients.

Semen alterations in porcine rubulavirus-infected boars are related to viral excretion and have implications for artificial insemination.

Porcine rubulavirus (PoRV), also known as blue eye disease (BED) of swine, causes respiratory and reproductive problems in pigs at several developmental stages. To study the effect of PoRV infection on semen production, five boars were infected with 1 x 10(6) TCID(50)/ml of PoRV strain PAC-3 and evaluated for 59 days post inoculation (DPI). Infected boars developed reproductive tract pathology that included swelling of the testes and epididymides. Analysis of the semen showed that the infection had little effect on semen production in four animals, but semen from one boar showed severe alterations in sperm concentration, motility, and morphology. When motility was analyzed in BTS-diluted semen after 24, 48, or 72 h, alterations were detected in all boars. Furthermore, viral antigen was detected in semen, the seminal plasma fraction, or sperm fraction from all boars. These results showed that PoRV is excreted via semen and, therefore, artificial insemination is a potential route of dissemination.

Porcine circovirus type 2 antibody detection in backyard pigs from Mexico City.

PCV2 antibodies have been found in pigs from all continents. However, this finding has been mainly studied in domestic swine reared under intensive production conditions. Mexico City, with a human population over 19 million in 2005, has both urban and rural areas. The pig production in its rural area is based on small family backyard farms. Taking into account this rather unique form of rearing pigs, the objective of this study was to determine the seroprevalence in backyard pigs from the rural area of Mexico City. A total of 695 backyard pig serum samples from 108 small family farms belonging to seven municipal areas were studied by immunoperoxidase monolayer assay technique. One hundred six out of the 108 family farms (98.14%) had at least one positive serum sample. On the other hand, 136 (19.57%), 264 (37.99%) and 248 (34.82%) pigs had low, intermediate and high titres to PCV2, respectively. Only 53 samples (7.63%) were negative for PCV2 antibodies. No apparent differences in antibody titre groups were observed among backyard pigs from the different municipal areas. In conclusion, the present study, the first one performed in this kind of extensively produced pigs, indicates that PCV2 is ubiquitous in backyard pigs from Mexico City.

[Transitional cell tumor in a 12-year-old patient]. / Urotelioma vesical en paciente de 12 años.

Transitional cell papillomas, which are tumors of epithelial origin, are the most common urinary tract neoplasm in adults but are extremely rare in children. In both groups the main location is the bladder, often around one of the ureteric orifices. Symptoms vary widely but the most frequent is gross hematuria. Transitional cell papillomas are histologically low-grade tumors and total excision is considered curative. However the rate of recurrence in the literature is high, and there is a potential for a more aggressive type of tumor. We report the case of a 12-year-old girl who presented with a transitional cell papilloma near the right ureteric orifice and describe the etiopathogenesis, diagnosis, treatment and outcome of this entity.

Integral pharmacological management of bone mineral disorders in chronic kidney disease (part II): from treatment of phosphate imbalance to control of PTH and prevention of progression of cardiovascular calcification.

INTRODUCTION: Chronic kidney disease-mineral and bone disorders (CKD-MBD) are associated with costly complications and dismal hard-outcomes. AREAS COVERED: In two comprehensive articles we review contemporary and future pharmacological options for treatment of phosphate (P) imbalance (part 1) and hyperparathyroidism (this part 2), taking into account CKD-accelerated cardiovascular calcification (CVC) processes. EXPERT OPINION: Improvements in CKD-MBD require an integral approach, addressing all three components of the CKD-MBD triad. Here, initial guidance to control hyperparathyroidism is provided, taking into account the presence/absence of CVC. We include also measures for patients at risk of adynamic bone disease or suffering from calciphylaxis. Many epidemiological studies (relating to vitamin D) and thorough analyses of recent randomized clinical trials (of cinacalcet) point towards benefits of attempting to improve biochemical parameters while trying to, at least, avoid progression of CVC by more rational use of intestinal P-binders and low-dose vitamin D derivatives and/or calcimimetics. This approach does not seem to be far away from significantly improving hard-outcomes, at least in the dialysis population. The availability of new drugs and the performance of randomized clinical trials should ultimately lead to define earlier, clearer, and more cost-effective patient stratification and biochemical targets with consequent significant clinical improvements.

Integral pharmacological management of bone mineral disorders in chronic kidney disease (part I): from treatment of phosphate imbalance to control of PTH and prevention of progression of cardiovascular calcification.

INTRODUCTION: Chronic kidney disease-mineral and bone disorders (CKD-MBD), involving a triad of laboratory and bone abnormalities, and tissue calcifications, are associated with dismal hard-outcomes. AREAS COVERED: In two comprehensive articles, we review contemporary and future pharmacological options for treatment of phosphate (P) imbalance (this part 1) and hyperparathyroidism (part 2), taking into account CKD-accelerated atheromatosis/atherosclerosis and/or cardiovascular calcification (CVC) processes. EXPERT OPINION: Improvements in CKD-MBD require an integral approach, addressing all three components of the CKD-MBD triad. Individualization of treatment with P-binders and combinations of anti-parathyroid agents may improve biochemical control with lower incidence of undesirable effects. Isolated biochemical parameters do not accurately reflect calcium or P load or bone activity and do not stratify high cardiovascular risk patients with CKD. Initial guidance is provided on reasonable therapeutic strategies which consider the presence of CVC. This part reflects that although there is not an absolute evidence, many studies point to the need to improve P imbalance while trying to, at least, avoid progression of CVC by restriction of Ca-based P-binders if economically feasible. The availability of new drugs (i.e. inhibitors of intestinal transporters), and studies including early CKD should ultimately lead to clearer and more cost/effective clinical targets for CKD-MBD.

Inhibition of T cell mitogenesis by nitrofurans.

A group of nitrofurans (5-nitro-2-furaldehyde, nifuroxime, nitrofurazone, nitrofurantoin, 5-nitro-2-furoic acid and 2-nitrofuran) were evaluated for inhibition of mitogenesis (DNA synthesis) in human peripheral blood T cells. T cells, either triggered by phorbol myristate acetate (PMA) or in the presence of accessory cells, were activated with a specified mitogen [phytohemagglutin (PHA), concanavalin A (ConA), or anti-CD3] and the amount of tritiated thymidine incorporated into DNA was determined. The results obtained indicate that nitrofurans inhibit mitogenesis irrespective of activator. 5-Nitro-2-furaldehyde was much more inhibitory than the other compounds, while 2-nitrofuran was less inhibitory. When the aldehyde group (5-nitro-2-furaldehyde) was replaced by a carboxyl group (5-nitro-2-furoic acid), the inhibitory activity was also reduced greatly. These results show that while the nitro group alone confers inhibitory activity to the furan ring, the group at the 2 position is crucial. In general, the mitogenic response of purified T cells (lacking accessory cells) triggered by PMA (phorbol ester) was inhibited less than that of the T cell-accessory cell system. With the latter, 50% inhibition of T cell mitogenesis was achieved by nifuroxime, nitrofurazone, and nitrofurantoin at 45-51 and 34-39 microM with PHA and ConA respectively. When purified T cells were used, the values were 71-85 and 55-60 microM respectively. For a given drug concentration, mitogenesis was more inhibited when induced by ConA or anti-CD3 than by PHA. The importance of using a single cell system (purified T cells) was emphasized by the interesting finding that only this system showed enhancement of mitogenesis, up to 35-40% at low drug levels. With the exception of the nitrofuraldehyde, the nitrofurans at strongly inhibitory levels were only moderately cytotoxic, exhibiting 62-85% cell survival after exposure to drug for 68 hr. Our results suggest that nitrofurans inhibit T cell mitogenesis by a relatively non-toxic mechanism; these results are comparable to those obtained for mammalian cells under aerobic conditions.

Serotonergic receptors in the brain of in utero undernourished rats.

In this study, we report that 5-HT(1A) receptors are already present in fractions of axonal growth cones, from the normal rat fetal brain (E-17). Also, in utero undernourished (UN) rat pups at birth show a noteworthy enhancement in the B(max) of [3H]5-hydroxytryptamine (5-HT) and [3H]8-hydroxy-(2-N,N-dipropilamin)-tetralin (([3H])8-OH-DPAT), in the brainstem and cerebral cortex up to the second week after birth. Afterwards, there is a significant decrease in the binding of these ligands. [125I]Cyanopindolo binding in the cerebral cortex only showed a decrease in the same period. An elevation of brain serotonin in both regions was also present. These findings together, suggest that the mechanisms of regulation of serotonergic receptors' expression during the period studied, may not depend on the amount of neurotransmitter in the synaptic cleft, because in the early UN brain it would be expected only a lower receptor's density due to the chronic serotonin increase. On this basis, we propose that developmental activation of brain serotonin biosynthesis observed in early UN animals may disrupt the mechanism regulating the expression of 5-HT receptors during development.

Comparative evaluation of the CD4+CD8+ and CD4+CD8- lymphocytes in the immune response to porcine rubulavirus.

The porcine immune system is unique in the expression of CD4+CD8+ (double-positive, DP) lymphocytes. These cells have been associated with immunological memory due to their gradual increase with age, the expression of memory phenotype and their ability to respond to recall viral antigen. This work analyzes the biological function of CD4+CD8- and CD4+CD8+ lymphocytes in the immune response to porcine rubulavirus (PRv). CD4+CD8- cells isolated from pigs 3 weeks after infection with porcine rubulavirus proliferated in response to homologous virus and generated lymphoblasts which were predominantly of the CD4+CD8+ phenotype, whereas stimulation with mitogen induced proliferation but did not switch the phenotype. CD4+CD8- lymphocytes isolated after 10 weeks of infection proliferated in response to phytohemagglutinin (PHA) but did not proliferate in response to homologous virus and did not change their phenotype, whereas CD4+CD8+ lymphocytes proliferated in response to PHA and to viral antigen. The cytokine profile of both lymphocyte populations showed the presence of IL-2 and IL-10 transcripts, quantitation demonstrated that CD4+CD8+ cells expressed mainly IL-10, whereas CD4+CD8- lymphocytes expressed primarily IL-2. Our results show that CD4+CD8- lymphocytes in the early phase of porcine rubulavirus infection can be converted to double-positive cells expressing IL-10 in an antigen-dependent manner, and that CD4+CD8- T-cells late in infection do not acquire CD8.

The use of biotin-labeled cDNA probes for the detection of infectious bursal disease viruses.

A cDNA library was prepared from the double-stranded RNA genome of the infectious bursal disease virus (IBDV) strain ST-C. The cDNA molecules were annealed into the plasmid pUC9 and used to transform Escherichia coli strain JM107. A cDNA clone that contained IBDV-specific nucleotide sequences was selected and designated STC-1. Radiolabeled probes were prepared from STC-1 and hybridized to genome segment A of ST-C in a northern blot hybridization assay. The STC-1 cDNA was 448 base pairs in length, and its nucleotide sequence indicated that it is located near the VP-2/VP-4 junction in IBDV genome segment A. Biotin-labeled probes were prepared from STC-1 and used in a dot-blot hybridization assay to detect IBDV. Under relatively low stringency conditions of hybridization, the biotinylated probes detected four subtypes of IBDV serotype 1 and a serotype 2 IBDV isolate.

Sustained levels of ascorbic acid are toxic and immunosuppressive for human T cells.

Ascorbate (vitamin C) can protect from oxidative damage to DNA and lipids that may lead to aging, cancer, and other dysfunctions. However, we find that purified human T cells deteriorate if maintained in ascorbate in culture for 18 hrs. or more; viability and Il-2 synthesis are over 90% curtailed by ascorbate at 50 micrograms/ml. T cell proliferation and adhesion are severely suppressed at 10-25 micrograms/ml. Dihydro-ascorbate was much less toxic or suppressive. The suppressive effect of ascorbate appears irreversible, since removal of ascorbate after 18 hrs. did not restore the mitogenic response. Although moderate dietary levels of ascorbate often reach 250-1000 mg or more daily and appear beneficial, our data caution against sustained megadoses of ascorbate for treatment of patients with AIDS and cancer.

Is social support associated with greater weight loss after bariatric surgery?: a systematic review.

Social support may be associated with increased weight loss after bariatric surgery. The objective of this article is to determine impact of post-operative support groups and other forms of social support on weight loss after bariatric surgery. MEDLINE search (1988-2009) was completed using MeSH terms including bariatric procedures and a spectrum of patient factors with potential relationship to weight loss outcomes. Of the 934 screened studies, 10 reported on social support and weight loss outcomes. Five studies reported on support groups and five studies reported on other forms of social support (such as perceived family support or number of confidants) and degree of post-operative weight loss (total n = 735 patients). All studies found a positive association between post-operative support groups and weight loss. One study found a positive association between marital status (being single) and weight loss, while three studies found a non-significant positive trend and one study was inconclusive. Support group attendance after bariatric surgery is associated with greater post-operative weight loss. Further research is necessary to determine the impact of other forms of social support. These factors should be addressed in prospective studies of weight loss following bariatric surgery, as they may represent ways to improve post-operative outcomes.

Eosinophilic enteritis in association with systemic lupus erythematosus.

Eosinophilic gastroenteritis (EGE) is an uncommon disease and has rarely been reported in association with connective tissue diseases as systemic lupus erythematosus. We report a 36-year-old woman who developed recurrent episodes of abdominal pain, nausea, vomiting and melena. Complete blood counts showed elevated eosinophil counts. Ultrasound and CT-scan images studies were significant for bowel wall thickening and ascites. The patient underwent an exploratory laparotomy with a mesenteric biopsy and appendectomy that showed eosinophil infiltration in the muscularis propria, establishing the diagnosis of EGE. The patient developed pleural effusions, with laboratory studies showing haemolytic anaemia, thrombocytopenia, positive antinuclear antibody and anticardiolipin antibodies. The patient was treated with high-dose systemic corticosteroid therapy, with successful resolution of symptoms. Three months later, she developed a new episode of abdominal pain defined as intestinal pseudo-obstruction that was resolved without complications.

Vasectomía sin bisturí: resultados de una cohorte de 309 pacientes / No-scalpel vasectomy

Introducción: La vasectomía es una técnica segura y efectiva para el control de la fertilidad masculina. A pesar de esto, en el mundo se realizan el doble de esterilizaciones femeninas respecto a vasectomías, lo cual es más acentuado en países en vías de desarrollo. Objetivos: El objetivo de este trabajo es presentar nuestra experiencia y resultados en pacientes sometidos a una vasectomía sin bisturí. Materiales y Métodos: Se incluyó un total de 309 pacientes sometidos a una vasectomía sin bisturí entre junio de 2009 y mayo de 2010. En cada caso se evaluó la edad, tiempo operatorio, espermiograma a los 3 y 6 meses (en caso de ser necesario) y la morbilidad perioperatoria. Resultados: 281 pacientes (91 por ciento) se controlaron con al menos un espermiograma. En 189 pacientes (67 por ciento) se evidenció azoospermia en el espermiograma a los 3 meses. En 81 pacientes (29 por ciento) se evidenció un recuento con < 100.000 espermatozoides 100 por ciento inmóviles. En 9 pacientes (3,2 por ciento) se necesitó un segundo espermiograma para obtener menos de 100.000 espermatozoides 100 por ciento inmóviles. Seis pacientes (2 por ciento) consultaron por complicaciones menores: orquialgia, epididimitos o hematoma del sitio operatorio. A 2 años de seguimiento, no se registraron embarazos. Conclusiones: La vasectomía sin bisturí es un método seguro y reproducible de anticoncepción masculina, presentando tasas de efectividad superiores al resto de los métodos anticonceptivos. Un espermiograma de control debe ser realizado a los 3 meses de realizado el procedimiento. La tasa de falla temprana se estima en 0,3 por ciento de los pacientes.

Introduction: Vasectomy is a safe and effective technique of male fertility control. Despite this, in the world are carried out more than double female sterilization in comparison with vasectomies, that is more pronounced in less developed countries. Aims: To present our experience and results in patients undergoing a no-scalpel vasectomy. Methods and Material: A total of 309 patients undergoing a no-scalpel vasectomy between June 2009 and May 2010 were included. For each case was record age, operative time, sperm count at 3 months post vasectomy and peri-operative morbidity. Results: 309 vasectomies were performed, 281 patients (91%) were controlled with at least one sperm count. Azoospermia was obtained in the first sperm count at 3 months in 189 patients (67%). In 81 patients (29%) were observed ≤ 100,000 sperm 100% immobile. 9 patients (3.2%) needed a second semen analysis and 2 patients a third one for less than 100,000 sperm that were 100% immobile. 6 patients (2%) consulted by minor complications such as postoperative pain, epididymitis or hematoma with spontaneous resolution. Conclusions: Vasectomy is a safe and reproducible method of male contraception, presenting an effectiveness rates higher than others contraceptive methods. There are no absolute contraindications for performing the procedure. A sperm count should be done at 3 months of the procedure. The early failure rate in our study is 0.3%.

Drug-drug interactions of non-steroidal anti-inflammatory drugs with other drugs in patients with rheumatic diseases.

OBJECTIVE: To determine the prevalence of and identify drug-drug interactions (DDI) between non-steroidal anti-inflammatory drugs (NSAID) and other drugs in a prescription database of patients with rheumatic iseases. MATERIAL AND METHODS: This is a cross-sectional study of a drug prescription database saving information on 35,000 individuals who benefited from a pre-paid medical system serving bank employees and their relatives. The analysis included one year period (from January to December 1998). NSAID-DDI were listed and classified into 3 levels (1: minor; 2: moderate, and 3: high health risk or death risk) according to DRUGDEX(r) as well as MEDLINE and EMBASE search. RESULTS: We analyzed 3,207 NSAID prescriptions (1.7±1.6 per patient) to 1,855 rheumatic patients (adults: 76.7%; geriatric: 20.2%, and pediatric: 3.0%; soft tissue rheumatism: 52%; osteoarthritis: 19%, and rheumatoid arthritis: 10%). There were 648 (20.20%) NSAID-DDI prescriptions: 594 (91.66%) corresponded to level 1; 46 (7.09%) to level 2, and 8 (1.23%) to level 3. In addition, 96 (2.99%) prescriptions included NSAID duplications. Interestingly, we found no NSAID-DDI with anticoagulants, anticonvulsants, and oral hypoglycemiants were found. CONCLUSIONS: The prevalence of NSAID-DDI prescriptions to 1,855 rheumatic patients was 20.20% in one year. NSAID-DDI was mostly (91.66%) level 1, and rarely (1.23%) level 3. NSAID duplications were found in 2.99%. These results provides information on the frequency of prescriptions with DDI, which might potentially produce harmful effects and data, which may help in the development of studies searching for the clinical relevance of NSAID-DDI.