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Differentiation of patients with rheumatic fever from those with inactive rheumatic heart disease using the artificial subcutaneous nodule test, myocardial reactive antibodies, serum immunoglobulin and serum complement levels.

Thirty patients with acute rheumatic fever, and 20 with inactive rheumatic heart disease were studied in order to determine parameters which differentiate the two groups. Subcutaneous nodules developed following the subcutaneous injection of autologous blood in 16 of 26 patients with rheumatic fever but in none of the 20 with inactive rheumatic heart disease (P = 0.00001). Myocardial reactive antibodies were found in 14 of 30 patients with rheumatic fever, and in 4 of 20 with inactive rheumatic heart disease (P = 0.0505). Significantly higher values of IgG (P less than 0.001), IgA (P less than 0.01), C3 (P less than 0.001) and C4 (P less than 0.001) were observed in rheumatic fever patients. There was no significant difference in IgM levels. A combination of three variables (artificial subcutaneous nodules, IgA, and C3) had a sensitivity of 87% and a specificity of 100% for rheumatic fever. A combination of two variables (artificial subcutaneous nodules and IgA) had a sensitivity of 84% and a specificity of 100% for rheumatic fever.

Adolescent↗

Characterization of Nops, nodulation outer proteins, secreted via the type III secretion system of NGR234.

The nitrogen-fixing symbiotic bacterium Rhizobium species NGR234 secretes, via a type III secretion system (TTSS), proteins called Nops (nodulation outer proteins). Abolition of TTSS-dependent protein secretion has either no effect or leads to a change in the number of nodules on selected plants. More dramatically, Nops impair nodule development on Crotalaria juncea roots, resulting in the formation of nonfixing pseudonodules. A double mutation of nopX and nopL, which code for two previously identified secreted proteins, leads to a phenotype on Pachyrhizus tuberosus differing from that of a mutant in which the TTSS is not functional. Use of antibodies and a modification of the purification protocol revealed that NGR234 secretes additional proteins in a TTSS-dependent manner. One of them was identified as NopA, a small 7-kDa protein. Single mutations in nopX and nopL were also generated to assess the involvement of each Nop in protein secretion and nodule formation. Mutation of nopX had little effect on NopL and NopA secretion but greatly affected the interaction of NGR234 with many plant hosts tested. NopL was not necessary for the secretion of any Nops but was required for efficient nodulation of some plant species. NopL may thus act as an effector protein whose recognition is dependent upon the hosts' genetic background.

Amino Acid Sequence↗

NolL of Rhizobium sp. strain NGR234 is required for O-acetyltransferase activity.

Following (iso)flavonoid induction, nodulation genes of the symbiotic nitrogen-fixing bacterium Rhizobium sp. strain NGR234 elaborate a large family of lipooligosaccharidic Nod factors (NodNGR factors). When secreted into the rhizosphere of compatible legumes, these signal molecules initiate root hair deformation and nodule development. The nonreducing glucosamine residue of NodNGR factors are N acylated, N methylated, and mono- or biscarbamoylated, while position C-6 of the reducing extremity is fucosylated. This fucose residue is normally 2-O methylated and either sulfated or acetylated. Here we present an analysis of all acetylated NodNGR factors, which clearly shows that the acetate group may occupy position C-3 or C-4 of the fucose moiety. Disruption of the flavonoid-inducible nolL gene, which is preceded by a nod box, results in the synthesis of NodNGR factors that lack the 3-O- or 4-O-acetate groups. Interestingly, the nodulation capacity of the mutant NGROmeganolL is not impaired, whereas introduction of the nod box::nolL construct into the related strain Rhizobium fredii USDA257 extends the host range of this bacterium to Calopogonium caeruleum, Leucaena leucocephala, and Lotus halophilus. Nod factors produced by a USDA257(pnolL) transconjugant were also acetylated. The nod box::nolL construct was also introduced into ANU265 (NGR234 cured of its symbiotic plasmid), along with extra copies of the nodD1 gene. When permeabilized, these cells possessed acetyltransferase activity, although crude extracts did not.

Acetylation↗

Thyroid nodules after high-dose external radiotherapy. Fine-needle aspiration cytology in diagnosis and management.

From 1974 to 1980, we studied six patients, five treated for Hodgkin's disease and one for a hemangioma, in whom thyroid nodules developed after high-dose (2,500 to 4,000 rad) thyroid radiation. Fine-needle aspiration cytology of the thyroid in five cases showed moderate to severe cellular atypia in four (three of whom had elevated serum thyrotropin levels) and mild atypia in one (who was overtly hypothyroid). In the four with marked atypia, surgical pathology disclosed Hashimoto's thyroiditis in one, diffuse hyperplasia and epithelial atypia in two, and a sclerosing papillary carcinoma in one. Among five patients who had total thyroidectomy, three had carcinoma (microscopic papillary, papillary-follicular, or sclerosing papillary) in addition to one or more follicular adenomas or colloid nodules. Thyroid nodules and neoplasia may be more common than previously appreciated in patients exposed to high-dose radiation.

Adenocarcinoma↗

Endoreduplication and activation of the anaphase-promoting complex during symbiotic cell development.

Postembryonic development of plant organs requires a constant interplay between the cell cycle and the developmental programs. Upon endo- and exogenous signals, plant cells can enter, exit or modify the cell cycle. Alteration of mitotic cycles to endoreduplication cycles, where the genome is duplicated without mitosis, is common in plants and may play a role in cell differentiation. The switch from the mitotic to endocycles is regulated by Ccs52A, a plant orthologue of the yeast and animal Cdhl proteins, acting as substrate-specific activator of the anaphase-promoting complex E3 ubiquitin ligase. Here, several aspects of endoreduplication are discussed with special attention on nitrogen-fixing nodule development where endoreduplication is an integral part of symbiotic cell differentiation.

Anaphase-Promoting Complex-Cyclosome↗

The multilayer technique: A new and fast approach for flashlamp-pumped pulsed (FLPP) dye laser treatment of port-wine stains (preliminary reports).

BACKGROUND: The 585 nm flashlamp-pumped pulsed (FLPP) dye laser is an effective and established treatment for port-wine stains (PWSs) during childhood. Unfortunately, PWSs tend to darken in color and may thicken or develop nodules as the lesions age, thereby making treatment difficult in adult patients since they may require several laser sessions producing unpredictable results. The aim of this article is to present and discuss the results obtained in four adult patients with PWS by use of a new approach in FLPP dye laser treatment. OBJECTIVE: The goal of this technique was to damage, during the same treatment, the lesions at both deep and superficial levels and to reduce the number of laser sessions required to obtain the most effective eradication of hypertrophied PWSs. METHODS: Four patients (two men and two women aged 54, 57, 49, and 61, respectively) were referred for treatment of congenital hypertrophied PWS of the face. Every dye laser session consisted of two laser passes. During the first pass the wavelength ranged from 590 to 600 nm with a long pulse (1.500 microseconds), while the second pass was performed utilizing the classic short pulse (450 microseconds) and wavelength (585 nm). Successive treatments were performed at 6- to 8-week intervals. RESULTS: All four patients had a complete clearing of their PWS after a number of treatments, ranging from three to five sessions. Three of them (one man and the two women) experienced extremely mild blistering in a limited small area that healed in approximately 10 days without scarring. The laser sessions were well tolerated by all patients. None of the patients developed atrophic or hypertrophied scars or dyschromia. CONCLUSIONS: Our results show an excellent response in all patients with just a few treatments (three to five sessions) and, in spite of two passes, only mild side effects that are probably limited due to cooling of the skin. We also observed a flattening and reduction of the nodules.

Female↗

The mobility outcome for amputees with rheumatoid arthritis is poor.

This study examines the reasons for amputation of the lower limb in 11 patients with RA. The reasons for amputation were ischaemia due to peripheral vascular disease; rheumatoid vasculitis; or complications of knee arthroplasty. Delayed healing was common in patients on steroid therapy. The patients had difficulty donning and doffing their prostheses due to poor hand function and impaired agility. Walking training and rehabilitation were slow and tended to precipitate a generalized flare-up of disease activity. Rheumatoid nodules developed on pressure points between stump and socket. Despite these problems the majority of patients preferred to wear a prosthesis for help in transferring in and out of the wheelchair, to complete their appearance and for limited mobility. Most found using an electric powered indoor wheelchair helped maintain their independence.

Adult↗

The role of ultrasonography-guided fine-needle aspiration biopsy in the management of nonpalpable and palpable thyroid nodules.

The introduction of highly sensitive imaging techniques has made it possible to detect many nonpalpable thyroid nodules (non-PTN). We investigated the value of ultrasound-guided fine-needle aspiration biopsy (US-guided FNAB) as a diagnostic tool in the management of non-PTN as well as palpable thyroid nodules (PTN) that were considered difficult to aspirate without guidance. US-guided FNAB was performed on a total of 119 nodules (71 palpable and 48 nonpalpable) from 119 patients between 1992 and 1996. All available clinical and follow-up data were reviewed. Surgical follow-up was available in 24 cases. The patients included 100 females and 19 males ranging in age from 9 to 81 years (average, 51 years). FNA diagnoses (PTN versus non-PTN) included papillary carcinoma (12.7% [9/71] versus 4.2% [2/48], follicular neoplasm (16.9% [12/71] versus 0%), medullary carcinoma (1.4% [1/71] versus 0%), atypical cytology (5.6% [4/71] versus 2.1% [1/48], non-neoplastic thyroid (63.4% [45/71] versus 85.4% [41/48]) and unsatisfactory (0% versus 8.3% [4/48]). In 2 cases of occult papillary carcinoma, risk factors included radiation exposure (1 case) and a newly developed nodule during follow-up for hypothyroidism (1 case). Subsequent surgical follow-up (24 cases) confirmed the FNA findings, except for a case of Hürthle cell adenoma and 1 of Hashimoto's thyroiditis diagnosed as papillary carcinoma and follicular neoplasm, respectively. US-guided FNAB in most non-PTN are diagnosed as benign. For most patients with non-PTN and without any high-risk factors, a conservative approach such as clinical follow-up may be a more cost effective and logical approach. In contrast, US-guided FNAB is more useful in diagnosing biologically significant lesions in PTN that may be difficult to aspirate without guidance.

Adolescent↗

The expression of MaEXP1, a Melilotus alba expansin gene, is upregulated during the sweetclover-Sinorhizobium meliloti interaction.

Expansins are a highly conserved group of cell wall-localized proteins that appear to mediate changes in cell wall plasticity during cell expansion or differentiation. The accumulation of expansin protein or the mRNA for specific expansin gene family members has been correlated with the growth of various plant organs. Because expansin proteins are closely associated with plant cell wall expansion, and as part of a larger study to determine the role of different gene products in the legume-Rhizobium spp. symbiosis, we investigated whether a Melilotus alba (white sweetclover) expansin gene is expressed during nodule development. A cDNA fragment encoding an expansin gene (EXP) was isolated from Sinorhizobium meliloti-inoculated sweetclover root RNA by reverse-transcriptase polymerase chain reaction using degenerate primers, and a full-length sweetclover expansin sequence (MaEXP1) was obtained using 5' and 3' rapid amplification of cDNA end cloning. The predicted amino acid of the sweetclover expansin is highly conserved with the various alpha-expansins in the GenBank database. MaEXP1 contains a series of eight cysteines and four tryptophans that are conserved in the alpha-expansin protein family. Northern analysis and whole-mount in situ hybridization analyses indicate that MaEXP1 mRNA expression is enhanced in roots within hours after inoculation with S. meliloti and in nodules. Western and immunolocalization studies using a cucumber expansin antibody demonstrated that a cross-reacting protein accumulated in the expanding cells of the nodule.

Amino Acid Sequence↗

Differential display analysis of RNA accumulation in arbuscular mycorrhiza of pea and isolation of a novel symbiosis-regulated plant gene.

Differential RNA display was used to analyze gene expression during the early steps of mycorrhiza development on Pisum sativum following inoculation with Glomus mosseae. Seven out of 118 differentially displayed cDNA fragments were subcloned and sequenced. One fragment corresponded to part of the fungal 25S ribosomal RNA gene and a second one showed similarity to a human Alu element. The others were derived from plant genes of unknown function. One of the fragments was used for the isolation of a full-length cDNA clone. It corresponded to a single-copy gene (psam1) which is induced during early symbiotic interactions, and codes for a putative transmembrane protein. Northern and RNA dot blot analyses revealed enhanced accumulation of psam1 RNA after inoculation with G. mosseae of wild-type pea and an isogenic mutant deficient for nodule development (Nod-, Myc+).

Base Sequence↗

Leiomyomatosis peritonealis disseminata in association with Currarino syndrome?

BACKGROUND: Leiomyomatosis peritonealis disseminata (LPD) is a rare disease in which multiple smooth muscle or smooth muscle-like nodules develop subperitoneally in any part of the abdominal cavity. No reports of multiple congenital malformations associated with LPD have been found in the English literature. CASE PRESENTATION: A 27 year-old patient referred to our gynaecology unit for pelvic pain, amenorrhoea, stress incontinence, chronic constipation and recurrent intestinal and urinary infections. Multiple congenital malformations had previously been diagnosed. Most of these had required surgical treatment in her early life: anorectal malformation with rectovestibular fistula, ectopic right ureteral orifice, megadolichoureter and hemisacrum. An ultrasound scan and computed tomography performed in our department showed an irregular, polylobate, complex 20 cm mass originating from the right pelvis that reached the right hypochondrium and the epigastrium. The patient underwent laparotomy. The three largest abdominal-pelvic masses and multiple independent nodules within the peritoneum were progressively removed. The histological diagnosis was of LPD. CONCLUSION: The case we report is distinctive in that a rare acquired disease, LPD, coexists with multiple congenital malformations recalling a particular subgroup of caudal regression syndrome: the Currarino syndrome.

Abnormalities, Multiple↗

Recurrence of thyroid nodules after surgical removal in patients irradiated in childhood for benign conditions.

To determine the incidence of benign thyroid nodules and the risk factors for their recurrence after surgical removal, we followed 511 patients for 1 to 40.6 years (median, 11.2) after surgery for benign thyroid nodules arising after local irradiation for unrelated benign diseases in childhood. Recurrent thyroid nodules developed in 100 patients (19.5 percent). The risk of recurrence correlated inversely with the amount of thyroid tissue removed. Women had a higher recurrence rate than men (28.4 percent vs. 10.3 percent; P less than 0.05). Among the 299 patients who had been treated with thyroid hormone at the discretion of their physicians to suppress thyroid-stimulating hormone, 25 had recurrences (8.4 percent), as compared with 72 of 201 patients who did not receive thyroid hormone (35.8 percent) (hazard ratio taking into account the extent of surgery and the patient's sex, 2.5; 95 percent confidence interval, 1.5 to 4.1). Histologic analysis of the 73 tissue samples from patients with recurrences showed that 14 samples (19.2 percent) were malignant. Thyroid hormone treatment had no effect on the rate of thyroid cancer. We conclude that radiation-associated benign thyroid nodules have a high recurrence rate, similar to that reported among nonirradiated patients with benign thyroid nodules. We also conclude that treatment with thyroid hormone decreases the risk of benign recurrences, particularly in women, but not the risk of cancer.

Adult↗

Seeding of gallbladder carcinoma along the tract after percutaneous transhepatic choledochoscopy.

An implant metastasis developed along the tract after percutaneous transhepatic cholecystoscopy 5 months after initial placement of the catheter into the gallbladder. Percutaneous transhepatic cholecystoscopy is conducted by insertion of choledochofiberscope into the gallbladder through the dilated sinus tract established around a catheter inserted into the gallbladder. The catheter had been placed for total of 15 days. Two days after the removal of the catheter, cholecystectomy supplemented by partial resection of the liver was carried out because of the extent of a gallbladder carcinoma. The choledochofiberscope used was Olympus CHF type 4B, with a thinner flexible part 5.0 mm in diameter. It is not surprising that carcinoma of the biliary tract or pancreas may seed along the tract of a biliary drainage catheter, but only few reports describing such incidence related to percutaneous transhepatic biliary drainage have previously been published. Physicians should be aware of this complication whenever a firm nodule develops at the site of previous entry of a biliary drainage catheter in a patient with malignant obstruction.

Adenocarcinoma↗

Identification of glyA as a symbiotically essential gene in Bradyrhizobium japonicum.

A Bradyrhizobium japonicum Tn5 mutant (strain 3160) induced numerous, tiny, white nodules which were dispersed over the whole root system of its natural host plant, soybean (Glycine max). These ineffective, nitrogen non-fixing pseudonodules were disturbed at a very early step of bacteroid and nodule development. Subsequent cloning and sequencing of the DNA region mutated in strain 3160 revealed that the Tn5 insertion mapped in a gene that had 60% homology to the Escherichia coli glyA gene coding for serine hydroxymethyltransferase (SHMT; E.C.2.1.2.1.). SHMT catalyses the biosynthesis of glycine from serine and the transfer of a one-carbon unit to tetrahydrofolate. The B. japonicum glyA region was able to fully complement the glycine auxotrophy of an E. coli glyA deletion strain. Although the Tn5 insertion in B. japonicum mutant 3160 disrupted the glyA coding sequence, this strain was only a bradytroph (i.e. a leaky auxotroph). Thus, B. japonicum may have an additional pathway for glycine biosynthesis. Nevertheless, the glyA mutation was responsible for the drastic symbiotic phenotype visible on plants. It may be possible, therefore, that a sufficient supply with glycine and/or a functioning C1-metabolism are indispensable for the establishment of a fully effective, nitrogen-fixing root nodule symbiosis.

Amino Acid Sequence↗

Genetic analysis of Rhizobium meliloti bacA-phoA fusion results in identification of degP: two loci required for symbiosis are closely linked to degP.

The function of the Rhizobium meliloti bacA gene, which is a homolog of the Escherichia coli sbmA gene, is required for an intermediate step in nodule development. A strain carrying the bacA386::TnphoA fusion was mutagenized with N-methyl-N'-nitro-N-nitrosoguanidine, and three mutants that had higher levels of alkaline phosphatase activity were identified. The mutations in these strains were recessive and mapped to the same genetic locus. The gene affected by these mutations was identified and sequenced and was found to be a homolog of the E. coli degP gene, which encodes a periplasmic endopeptidase. Although degP function is important for the virulence of certain intracellular pathogens of mammals, it is not required for the R. meliloti-alfalfa symbiosis. The genetic analyses involving degP were complicated by the presence of a locus immediately upstream of depP that was lethal when present in multiple copies in a DegP- background. R. meliloti derivatives carrying insertion mutations in this locus displayed an N,N,N',N'-tetramethyl-p-phenylenediamine oxidase-negative phenotype, elicited the formation of white cylindrical nodules that did not fix nitrogen, and grew slowly in rich medium, suggesting that the locus was a cyc gene encoding a protein involved in the biosynthesis of a component or components of a respiratory chain. The previously identified fix-382::TnphoA, which similarly causes the formation of white cylindrical nodules that do not fix nitrogen, was shown to affect a gene that is separate from this cyc gene but extremely closely linked to it.

Alkaline Phosphatase↗

Six nodulation genes of nod box locus 4 in Rhizobium meliloti are involved in nodulation signal production: nodM codes for D-glucosamine synthetase.

The nucleotide sequence of the nod box locus n4 in Rhizobium meliloti was determined and revealed six genes organized in a single transcriptional unit, which are induced in response to a plant signal such as luteolin. Mutations in these genes influence the early steps of nodule development on Medicago, but have no detectable effect on Melilotus, another host for R. meliloti. Based on sequence homology, the first open reading frame (ORF) corresponds to the nodM gene and the last to the nodN gene of Rhizobium leguminosarum. The others do not exhibit similarity to any genes sequenced so far, so we designated them as nolF, nolG, nolH and nolI, respectively. We found that the n4 locus, and especially the nodM and nodN genes, are involved in the production of the root hair deformation (Had) factor. NodM exhibits homology to amidotransferases, primarily to the D-glucosamine synthetase encoded by the glmS gene of Escherichia coli. We demonstrated that in E. coli the regulatory gene nodD together with luteolin can activate nod genes. On this basis we showed that nodM complemented an E. coli glmS- mutation, indicating that nodM can be considered as a glmS gene under plant signal control. Moreover, exogenously supplied D-glucosamine restored nodulation of Medicago by nodM mutants. Our data suggest that in addition to the housekeeping glmS gene of R. melioti, nodM as a second glmS copy provides glucosamine in sufficient amounts for the synthesis of the Had factor.

Amino Acid Sequence↗

The broad bean nodulin VfENOD18 is a member of a novel family of plant proteins with homologies to the bacterial MJ0577 superfamily.

Full-length transcript sequences were isolated from broad bean root nodules, which encode a novel nodulin designated VfENOD18. The corresponding transcripts were detected in early and in late stages of nodule development and were localized exclusively in the nitrogen-fixing zone III. The VfENOD18 sequence is not only homologous to a number of ESTs from various mono- and dicotyledonous plants, but also to the ATP-binding protein MJ0577 from Methanococcus jannaschii and to a range of bacterial proteins that belong to the MJ0577 superfamily. Hence, VfENOD18 is a member of a ubiquitous family of plant proteins that might function as ATP-binding proteins or ATPases. On the genomic level, VfENOD18 genes can be divided into two groups on the basis of differences in their 5' UTRs. One group lacks the 5' UTR region including the ATG initiation codon, whereas the second group contained the complete 5' UTR region. Further upstream of this VfENOD18 gene, a retrotransposon sequence was identified. The -14/-964 VfENOD18 promoter fragment was devoid of complete organ-specific elements known from other nodulin gene promoters. Nevertheless, this region was able to mediate full promoter activity in the central region of transgenic Vicia hirsuta root nodules.

5' Untranslated Regions↗

Pneumoperitoneum impairs blood flow and augments tumor growth in the abdominal wall.

BACKGROUND: Despite several clinical and experimental studies, the mechanisms behind the development of port site metastases in laparoscopic surgery have remained largely unknown. The current study was designed to investigate the effect of pneumoperitoneum on blood flow in the abdominal wall and its possible effects on tumor growth at this site. METHODS: A total of 40 Wistar Fu rats had a laser Doppler probe placed on their left rectus muscle and a suspension of 50,000 adenocarcinoma cells was injected into their right rectus muscle. The experimental group (n = 20) was insufflated with air at 10 mmHg for 45 min while abdominal blood flow was registered before and during insufflation and after exsufflation. The control group (n = 20) was not insufflated but the blood flow was recorded in the same manner. After 9 days, all animals were killed and the occurrence of tumor was observed. The tumors were analyzed with respect to weight and volume. RESULTS: The insufflation caused an 82% reduction in blood flow in the experimental group (p < 0.001). No reduction in blood flow was registered in the control group. Tumor nodules developed significantly more often in the insufflated group (20/20) compared to the controls (14/20) (p = 0.016). Tumor weight (p = 0.003) and volume (p < 0.001) were significantly increased in the insufflated group. CONCLUSIONS: Pneumoperitoneum seems to enhance tumor growth. It also causes a significant reduction in blood flow in the abdominal wall, which may contribute to the increased susceptibility of tumor take.

Adenocarcinoma↗