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Blood flow limitations of solute transport across the visceral peritoneum.

In a previous study, no limitations to urea transfer across the parietal peritoneum were demonstrated with decreases in local blood flow of 70%. It was hypothesized that the visceral peritoneum would have similar characteristics. To address this problem at the tissue level, diffusion chambers were affixed to the serosal side of the stomach, cecum, or liver of anesthetized rats (n = 6 each tissue), and solutions containing 14C urea were placed in the chamber. During each experiment, the local chamber blood flow was measured with laser Doppler flowmetry, and, simultaneously, the disappearance of the tracer versus time was determined under three conditions: control, after 60 to 70% blood flow reduction, and postmortem (flow = 0). The results showed no difference in the urea mass transfer coefficient (MTC; mean +/- SEM; cm/min x 10[3]) between control and blood flow reduction for the stomach (4.0 +/- 0.4 versus 3.6 +/- 0.3) or for the cecum (4.6 +/- 0.3 versus 4.0 +/- 0.3). However, the MTC was significantly decreased by local blood flow reduction in the liver (5.4 +/- 0.2 versus 2.6 +/- 0.2). Postmortem data demonstrated significant reductions in the MTC with blood flow equal to zero. It is concluded that a 60 to 70% blood flow reduction from control values does not limit solute transperitoneal transfer in the hollow viscera but causes significant changes in the mass transfer across the liver surface. Because the liver makes up only a small portion of the effective exchange area, overall transperitoneal solute transfer should not be greatly affected by significant decreases in blood flow.

Animals↗

Natural and acquired macrolide resistance in mycobacteria.

The genus Mycobacterium contains two of the most important human pathogens, Mycobacterium tuberculosis and Mycobacterium leprae, the etiologic agents of tuberculosis and leprosy, respectively. Other mycobacteria are mostly saprophytic organisms, living in soil and water, but some of them can cause opportunistic infections. The increasing incidence of tuberculosis as well as infections with non-tuberculous mycobacteria (NTM) in AIDS patients has renewed interest in molecular mechanisms of drug resistance in these pathogens. Mycobacteria show a high degree of intrinsic resistance to most common antibiotics. For instance, species from the M. tuberculosis complex (MTC) are intrinsically resistant to macrolides. Nevertheless, some semi-synthetic macrolides as the erythromycin derivatives clarithromycin, azithromycin and most recently the ketolides, are active against NTM, particularly Mycobacterium avium, and some of them are widely used for infection treatment. However, shortly after the introduction of these new drugs, resistant strains appeared due to mutations in the macrolide target, the ribosome. The mycobacterial cell wall with its specific composition and structure is considered to be a major factor in promoting the natural resistance of mycobacteria to various antibiotics. However, to explain the difference in macrolide sensitivity between the MTC and NTM, the synergistic contribution of a specific resistance mechanism might be required, in addition to possible differences in cell wall permeability. This mini-review summarizes the current knowledge on the natural and acquired macrolide resistance in mycobacteria, gives an overview of potential mechanisms implicated in the intrinsic resistance and brings recent data concerning a macrolide resistance determinant in the MTC.

Anti-Bacterial Agents↗

Preserved prolactin fluctuations and response to metoclopramide in ovulatory, infertile, hyperprolactinemic women.

The study included 18 infertile, hyperprolactinemic women with preserved menstrual cycles. Among 13 women with consistently elevated prolactin (PRL) levels, 6 had either anovulatory cycles or luteal insufficiency, whereas 7 had apparently normal ovulatory cycles. Comparisons of these two groups showed that basal levels were similar but the PRL response to metoclopramide (MTC) and the day-to-day fluctuations were significantly lower in the group with altered ovulatory function. In this group the PRL response to MTC was significantly greater than in control patients with hyperprolactinemic amenorrhea (n = 12), but lower than in normal women (n = 10). The PRL response to MTC in hyperprolactinemic women with normal ovulatory function did not differ significantly from that of normal women, irrespective of whether hyperprolactinemia was sustained (n = 7) or intermittent (n = 5). The study indicates that the degree of autonomy of PRL secretion reflected by both the fluctuations in basal levels and the response to a dopamine antagonist, may be used to evaluate whether or not slightly elevated PRL levels are of clinical significance in relation to fertility.

Adult↗

Magnetic resonance imaging of the galactosemic dog eye using magnetization transfer contrast.

Magnetization transfer contrast (MTC) enhanced magnetic resonance (MR) imaging is a technique that generates high contrast images based on characteristic tissue differences resulting from the interaction of water and macromolecules. In this study, the feasibility of applying this technique to documenting the progression of osmotic sugar cataract formation was investigated in male beagles, initially 6 or 24 month old, fed a diet containing 30% galactose. MTC enhanced magnetic resonance imaging was periodically conducted on these animal's eyes at 2-Tesla. The lens MR images were compared to photographs obtained by photo-slit lamp and retroillumination photography. The MTC technique provided improved image details of the lens and anterior segment that documented osmotic changes from initial cortical vacuole formation to cortical and nuclear changes associated with advanced sugar cataracts. The latter could not be observed by photo-slit lamp or retroillumination photography.

Animals↗

Presumptive diagnosis of subclinical infections utilizing computer-assisted analysis of sequential enzyme-linked immunosorbent assays against multiple antigens.

One-hundred-seventy-two serum samples, collected sequentially from four flocks of egg- and meat-type chickens, were evaluated for antibodies to multiple infectious agents by enzyme-linked immunosorbent assay (MELISA). The MELISA system used provided simultaneous measurement of antibody titers against avian infectious bronchitis (IB), infectious bursal disease (BD), Newcastle disease, avian encephalomyelitis and reovirus infections, and Mycoplasma gallisepticum. The use of computer-generated graphic print outs of relative MELISA titers provided immediate visulization of over 740 data points and convenient detection of any temporal changes in median titer class (MTC). The temporally changing MTC, or flock profiles obtained, indicated that negligible or waning IB immunity may be a common occurrence in previously vaccinated commercial chickens. These profiles further suggested that, despite no IB revaccination, these same flocks experienced episodes of reexposure to IB which otherwise may have been difficult to detect by conventional clinical or diagnostic laboratory protocols. MELISA profiles and sequential histologic examinations of bursas of Fabricius also provided evidence of a possible BD vaccination problem in young chickens that also experienced excessive losses from coccidiosis, ulcerative enteritis, and Marek's disease. Short sampling intervals were found to foster the detection and definition of fluctuations in MTC which otherwise may have been missed.

Animals↗

A study of histopathological features of medullary carcinoma of the thyroid: cases from a single institute in India.

BACKGROUND: The microscopic features of medullary carcinoma have been described in world literature, together with its behavior and molecular biology. However, no large study has been reported from India. AIMS: This study aims to analyse the clinical, and especially the pathological features of medullary carcinoma of the thyroid, and the surrounding thyroid. MATERIALS AND METHODS: In this study a total of 234 cases of medullary thyroid carcinoma (MTC) were gathered over a period of 3 decades. The clinical presentation, the microscopic features and the clinical outcome were analyzed. RESULTS: MTC was found to be twice as common in men as in women and for some reason it occurred 10 years earlier in women. The histology revealed certain interesting features like the presence of apoptosis in over half of the tumors, in addition to the other common and not so common histological findings (encapsulated variant, small cell variants, follicular pattern, rosettes, oncocytic change, osteosarcoma-like pattern, and cribriform pattern). The adjacent thyroid in about 19% of the cases showed optically clear nuclei in the follicles that were close to the tumor cells. These features were similar to those seen in papillary thyroid carcinoma. CONCLUSIONS: The thyroid adjacent to MTC showed nuclear changes, which are also found in papillary carcinoma of the thyroid. The occasional concurrent occurrence of these two tumors and the involvement of the RET gene in both medullary and papillary carcinomas, makes this observation worth discussing and studying further.

Adolescent↗

RET germline mutation in codon 791 in a family representing 3 generations from age 5 to age 70 years: should thyroidectomy be performed?

OBJECTIVE: To describe a kindred with a rare RET germline mutation in codon 791 and discuss potential management strategies. METHODS: We present clinical and biochemical data as well as results of mutation analysis in our study subjects and provide an overview of related published reports. RESULTS: Multiple endocrine neoplasia type 2 (MEN 2) is a familial cancer syndrome characterized by the development of medullary thyroid carcinoma (MTC), pheochromocytoma, and parathyroid hyperplasia or adenoma. Germline mutations in RET are responsible for this autosomal dominant syndrome. Familial MTC is a variant of MEN 2A and can be caused by RET mutations in codon 791. Deaths from gene carriers with mutations in these codons have not yet been reported. In general, gene carriers with these RET mutations have late-onset MTC. Because only a few kindreds with this specific mutation have been identified and no long-term follow-up data are available, management of these patients can be a challenge. We illustrate the difficulties with decisions about not only when to perform thyroidectomy in these patients but also whether thyroidectomy should even be considered in such gene carriers with a benign course. Our reported kindred included four carriers with a codon 791 RET germline mutation, one of whom had the rare concomitant occurrence of acromegaly and MEN 2A. The 70-year-old mother had acromegaly and hyperparathyroidism but normal serum calcitonin levels and normal findings on thyroid ultrasound examination. She refused pentagastrin testing and any surgical intervention. The 37-year-old daughter had hypothyroidism, a small thyroid gland, and negative results of pentagastrin stimulation testing of calcitonin. The 18-year-old grandson also had a negative pentagastrin test result and normal thyroid ultrasound findings. The 5-year-old granddaughter had normal results of thyroid ultrasonography. In all patients, we recommended thyroidectomy. CONCLUSION: Prospective studies are needed to clarify which patients with codon 791 RET germline mutation should undergo thyroidectomy.

Acromegaly↗

Correlation of beta-actin messenger RNA localization with metastatic potential in rat adenocarcinoma cell lines.

The actin cytoskeleton is involved in the motility of tumor cells. It has been shown in several cell types that beta-actin mRNA is localized in the protrusions of cells in which actin is actively polymerized, and the ability to localize mRNA is correlated with the efficiency of motility. In this context, we studied the distribution of beta-actin mRNA in two different tumor cell lines and correlated it with their metastatic potential. The two cell lines used were the highly metastatic MTLn3 cells and nonmetastatic MTC cells. Nonmetastatic MTC cells have two different pools of beta-actin mRNA (perinuclear and at the leading edge), whereas highly metastatic MTLn3 cells have only a perinuclear distribution of beta-actin mRNA. These differences in mRNA localization are correlated with profound differences in the polarity and plasticity of cell motility of these cells in culture and the histopathology of primary breast tumors derived from these cells. In particular, MTLn3 cells are unpolarized by all cell shape and motility criteria in culture and in their histopathological organization in primary tumors. By comparison, MTC cells are polarized in all identical measurements. These results suggest that the increased plasticity of cell locomotion and the invasiveness of MTLn3 cells result from the failure of metastatic cells to localize beta-actin mRNA properly, causing them to be less polarized and therefore more flexible in their direction of motility. Thus, differences in the polarized organization of cells in the primary tumor that are correlated with beta-actin mRNA localization may have prognostic value in predicting metastatic potential.

Actins↗

[The experimental pathology and clinical treatment of microwave tissue coagulation for ocular superficial lesions].

OBJECTIVE: To investigate the therapeutic effect of a new therapy, microwave tissue coagulation (MTC), for treatment of ocular diseases. METHODS: HSE-5M Mircotaze (manufactured by HEIWA Electronic Industrial Co. Osaka, Japan) was used for the treatment of sixty-four cases with ocular superficial lesions. Animal experiment was performed with this technique before application in patients. Twelve rabbit monocular eyeballs were coagulated just posterior to the corneoscleral limbus in order to find a dose which is effective and safe for the human eye ball. The models of microwave coagulative lesions were made in the contralateral eyes of the twelve rabbit eyeballs. RESULTS: The coagulation-induced "ulcers" were exfoliated gradually and healed finally from three to seven days. Histopathology of the coagulative areas showed coagulative necrosis with infiltration of lymphocytes and plasma cells. The results of sixty-four cases treated with MTC technique show that the damaging effect of coagulation on tissues is very mild, so that it heals very soon and the clinical process is consistent with the experimental pathological process. CONCLUSIONS: If the dosage of the technique is controlled in the range of safety and effectiveness, the MTC therapy is effective for the treatment of superficial ocular disease. The clinical and pathological examinations demonstrate that the therapy does not damage the ocular structure.

Animals↗

Improved prediction of calcitonin normalization in medullary thyroid carcinoma patients by quantitative lymph node analysis.

BACKGROUND: The aim of this study was to identify better prognostic parameters for normalization of serum calcitonin in medullary thyroid carcinoma (MTC) patients. METHODS: In 73 patients who had undergone systematic lymph node dissection for MTC between September 1995 and November 1998, preoperative (n = 29) and postoperative (n = 65) basal and stimulated serum calcitonin were correlated with the pTNM classification and the number of positive regional lymph nodes and compartments. RESULTS: In contrast to pT and M, there was a significant correlation between postoperative calcitonin and the pN category. With rising numbers of positive lymph nodes (0, 1-9, 10-19, and > or = 20), postoperative basal and stimulated calcitonin increased exponentially, and gross distant metastases (M1) occurred more frequently (0%, 4%, 13%, and 50%; P = 0.013). Conversely, serum calcitonin was less often normalized (65%, 31%, 0%, and 0%; P = 0. 003). There was a close correlation between the number of positive lymph nodes and the number of affected compartments (P < 0.001; r = 0.93). Irrespective of location, involvement of 10 or more lymph nodes and more than 2 compartments precluded normalization of serum calcitonin. CONCLUSIONS: Quantitative lymph node analysis of MTC improves prediction of calcitonin normalization. When more than two compartments are involved, normalization of serum calcitonin cannot be attained. Surgery should then be less extensive and more directed at preventing local complications.

Adult↗

Protein-losing enteropathy is associated with peritoneal functional abnormalities in peritoneal dialysis patients.

OBJECTIVE: To evaluate the relationship between acquired peritoneal transport disorders and the presence of protein-losing enteropathy (PLE), and their contribution to the protein malnutrition in peritoneal dialysis (PD) patients. PATIENTS AND METHODS: We studied 31 clinically stable PD patients that received a fat overload diet for 3 days. We measured intestinal absorption of fecal fat (normal < 6 g/24-hour stool) and nitrogen (normal < 2 g/24-hr stool), intestinal protein permeability [fecal clearance of alpha1-antitrypsin (Calpha1AT) (normal < 12 mL/24-hr stool)], and nutritional markers [normalized protein nitrogen appearance (nPNA), half-life medium-term proteins, and body mass index]. Peritoneal solute transport was measured by mass transfer coefficient (MTC), and water transport by peritoneal ultrafiltration (UF) capacity. To define protein maldigestion it was necessary to find high fecal nitrogen values with normal Calpha1AT; PLE was defined when both values were elevated. RESULTS: High fecal nitrogen (mean 2.1+/-1 g/24-hr stool) and fat (mean 5.8+/-3.6 g/24-hr stool) were found in 15 patients; 6 patients had high Calpha1AT levels (PLE). These 6 patients showed a worse nutritional status: lower albumin (3.57+/-0.57 g/dL vs 3.98+/-0.38 g/dL, p < 0.05) and transferrin (243+/-70 mg/dL vs 272+/-44.3 mg/dL, p < 0.05), as well as lower triglycerides (131.3+/-31.7 mg/dL vs 187+/-116 mg/dL, p< 0.05). Higher urea MTCs were found in 10 patients, normal in 7, and lower in 14. Higher creatinine MTCs were found in 8 patients, normal in 15, and lower in 8. Normal peritoneal UF capacity was found in 25 and lower in 6 patients. These 6 patients showed higher urea and creatinine MTCs and Calpha1AT. A positive linear correlation between Calpha1AT, urea MTC (r = 0.56, p < 0.01), and creatinine MTC (r = 0.46, p < 0.01) was found. A similar situation occurred between Calpha1AT, fecal fat (r = 0.45, p < 0.05), and fecal nitrogen (r = 0.43, p < 0.05). Thirteen patients with previous history of peritonitis showed higher Calpha1AT than those without peritonitis (10.2+/-8 mL/24-hr stool vs 5.2+/-4.4 mL/24-hr stool, p < 0.05). CONCLUSIONS: We confirm that protein and fat malabsorption, maldigestion, and PLE are present in some PD patients. Higher fecal Calpha1AT is associated with malnutrition and poorer showings of the viability markers of peritoneal membrane function.

Adult↗

Glucocorticoids differentially inhibit expression of the RET proto-oncogene.

The RET proto-oncogene encodes a receptor tyrosine kinase activated by the binding of factors from the glial cell line-derived neurotrophic factor (GDNF) family to receptor-alpha components such as GDNF family receptor alpha-1 (GFR alpha-1). Mutations within the sequence of the RET proto-oncogene are associated with multiple endocrine neoplasia type 2 (MEN 2), an inherited tumor syndrome characterized by the development of medullary thyroid carcinoma (MTC) and other neuroendocrine tumors. Despite Northern analysis showing that RET is expressed in the majority of MTCs, the factors regulating this expression are poorly understood. To address this issue we examined RET expression in response to glucocorticoids in the TT cell line, derived from a metastatic MTC. The synthetic glucocorticoid dexamethasone was found to reduce RET expression at both mRNA and protein levels. This effect was dose responsive and maximal at 24 h. The reduction in RET mRNA was shown to be specific to glucocorticoids and was also seen in a primary MTC culture. Nuclear run-on studies revealed the reduction in steady-state RNA to be due to a decrease in RET mRNA transcription and the effect was shown to be independent of new protein synthesis or RNA stability. Dexamethasone was also found to exert an inhibitory effect upon cell growth, suggesting a potential use for glucocorticoids in the treatment of medullary carcinoma and MEN 2.

Carcinoma, Medullary↗

Reactive oxygen species are critical for the growth and differentiation of medullary thyroid carcinoma cells.

Reactive oxygen species have recently been demonstrated to play a role in numerous cellular signal transduction pathways. Here we investigate the involvement of H2O2 in Raf-1-mediated differentiation in the human medullary thyroid carcinoma (MTC) cell line TT:deltaRaf-1:ER. Catalase, but not Cu/Zn superoxide dismutase, completely inhibited Raf-1-induced differentiation of beta-estradiol-treated TT: deltaRaf-1:ER. In addition, catalase treatment down-regulated RET expression at both the mRNA and protein levels and induced apoptosis in the parental TT cell line and uninduced TT:deltaRaf-1:ER human MTC cells. These results implicate H2O2 as a downstream mediator of c-Raf-1-induced differentiation and as a survival factor in MTC cells.

Carcinoma, Medullary↗

[Multiple endocrine neoplasia: a clinical model for applying molecular genetic techniques].

Multiple endocrine neoplasias (MEN) are syndromes inherited as autosomal dominant. The application of the techniques of molecular biology has made possible the identification of the genes causing MEN 1 and 2. The gene responsible for MEN 1 belongs to the family of tumor suppressor genes and encodes for a protein named MENIN whose function remains to be elucidated. The identification of mutant MEN 1 gene carriers who are at risk of developing this syndrome requires frequent biochemical screening for the development of endocrine tumors. MEN 2 is a consequence of mutations in the Ret proto-oncogene (c-Ret). This gene encodes for a tyrosine kinase receptor thought to play a role in the development of neural crest-derived tissue. Members of kindred with either MEN 2A or MEN 2B should be screened by direct DNA testing early in life for mutations in c-Ret. Those with the mutation should be advised to have thyroidectomy at five years of age in children with MEN 2A and earlier in children with MEN 2B. Some cases of sporadic MTC are actually MEN 2A or Familial MTC after c-Ret testing is done, therefore routine application of this test is recommended in all cases of apparent sporadic MTC.

Adult↗

[MEN 2A syndrome: report of a family].

A typical three generation-family with MEN 2A (Multiple Endocrine Neoplasia, Type 2A) is described. Two brothers and their uncle were affected with medullary thyroid carcinoma (MTC) and pheochromocytoma. The diagnosis of MTC was based on calcitonin determinations after pentagastrin stimulation. Pheochromocytoma was clinically silent in two cases. A family screening with pentagastrin test led to the diagnosis of C cell hyperplasia in a 8-year boy, who was then successfully operated on. The authors discuss the value of pentagastrin test and genetic testing in the early diagnosis of MTC associated with MEN 2A syndrome.

Adult↗

The effectiveness of combined oral lysine acetylsalicylate and metoclopramide (Migpriv) in the treatment of migraine attacks. Comparison with placebo and oral sumatriptan.

In two, double-blind, randomised, clinical, trials (RCTs), oral lysine acetylsalicylate (1620 mg, equivalent to 900 mg aspirin) combined with metoclopramide (10 mg) (LAS + MTC) was compared with placebo, and with oral sumatriptan (100 mg) in one of these RCTs. In both RCTs the LAS + MTC combination was superior to placebo with therapeutic gains (percentage relief after active treatment minus percentage relief after placebo) of 30% and 31% for the first treated attack. These therapeutic gains are in the same range as those found for 100 mg oral sumatriptan, and in the comparative RCT the LAS + MTC combination was quite comparable to 100 mg sumatriptan, with success rates for the first attack of 57% and 53%, respectively.

Anti-Inflammatory Agents, Non-Steroidal↗

Chromogranin A: an additional tumor marker for postoperative recurrence and metastases of medullary thyroid carcinomas?

In this study, plasma concentrations of chromogranin A, calcitonin and carcinoembryonic antigen (CEA) were measured in 40 healthy volunteers as well as in 129 patients with recurrences and/or metastases of neuroendocrine tumors and of medullary thyroid carcinomas (MTCs). A double antibody assay was employed using polyclonal rabbit antibodies to a C-terminal fragment of the protein for detection of human chromogranin A. Using ROC analysis, a cutoff at 22 U/l chromogranin A was calculated. In patients with neuroendocrine tumours, much higher serum concentrations of chromogranin A than for patients with MTC (80% vs. 46%) were measured. The following sensitivities were found: chromogranin A; 46%, calcitonin 100%, CEA 52%. Furthermore, the mean values of chromogranin A concentrations correlated with the tumour mass and/or number of metastases in MTC and neuroendocrine tumours. Evaluation of follow-up studies remains to be completed; however, preliminary results showed similarities regarding the behaviour of chromogranin A and calcitonin. Despite the findings of this study, the observations could not confirm chromogranin A as a reliable marker for metastazing or recurrent MTC.

Biomarkers, Tumor↗

Preliminary evaluation of influence of gemcitabine (Gemzar) on proliferation and neuroendocrine activity of human TT cell line: immunocytochemical investigations.

The choice treatment of medullary thyroid carcinoma (MTC) is total thyroidectomy. It is difficult to evaluate effectiveness of chemotherapy due to the rare incidence of MTC. Gemcitabine is a new drug of antimetabolite nucleoside group used in treatment of cancers since 1996. The aim of this study was to evaluate the influence of gemcitabine on proliferation and neuroendocrine activity of human TT cell line derived from MTC. The cells were exposed to gemcitabine in the concentration of 10, 25 and 50 microg/ml for 24 hours. Immunocytochemical examinations were carried out by the method of avidin-biotin peroxidase complex (ABC) according to Hsu et al. to detect calcitonin, chromogranin A, synaptophysin and neuron-specific enolase in TT cells. A concentration-dependent inhibitory influence of gemcitabine on proliferative activity of TT cells was observed. It was also shown that the immunostaining was reduced, especially in case of neuron-specific enolase. Only the reaction detecting calcitonin was enhanced in persistin

Antimetabolites, Antineoplastic↗