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Sinorhizobium fredii isolates can be specifically identified by a 260 bp fragment from the nolXWBTUV locus.

A pair of primers homologous to the nolXWBTUV locus generated a 260 bp fragment by PCR only in the presence of Sinorhizobium fredii template DNA of different quality. This resulted in a fast and accurate method for the identification of S. fredii either from pure DNA, whole bacterial cells or nodule extracts. By means of two PCR fragments, one specific for S. fredii (260-bp) and the other specific for Bradyrhizobium japonicum (RSalpha), we found that S. fredii strain SMH12 and B. japonicum E109 were equally efficient at developing nodules on soybean plants grown under controlled environmental conditions.

Chromosome Mapping↗

Heterologous protection by Leishmania donovani for Leishmania major infections in the vervet monkey model of the disease.

The study was aimed at analyzing immunological cross-reactivity between Leishmania major and Leishmania donovani and possible cross-protection between the two parasite species in the vervet monkey model of the disease. Nine vervet monkeys (Cercopithecus aethiops) from the institute animal colony were sued in the study. Five of the animals had been previously infected with L. donovani but had remained asymptomatic while the other four animals were naive and comprised the control group. Immunological responses to both L. major and L. donovani antigens in the five animals with prior exposure to L. donovani were examined before challenge. High antibody titers to the two antigens were demonstrated in an enzyme-linked immunosorbent assay, but the antibody titers to L. donovani were significantly higher than those to L. major (P < 0.005). Positive in vitro peripheral blood leucocyte (PBL) proliferation to L. major and L. donovani antigens was also demonstrated, but there was no significant difference in the response to the two antigens (P > 0.1). High and varying levels of interferon gamma (IFN-gamma) were secreted in PBL from the five vervet monkeys when stimulated with L. major antigen, but vervet monkey 1296 secreted marginal levels of IFN-gamma. When the animals were challenged intradermally with 1 x 10(5) virulent L. major promastigotes mixed with sandfly vector salivary gland lysate all four vervet monkeys in the control group developed nodules of varying sizes at the inoculation sites that eventually ulcerated. However, nodule formation and ulceration occurred at different times among these animals. The other five animals (animals with prior exposure to L. donovani) did not pick up the infection at all, but one animal from this group, vervet monkey 1296, developed a transient lesion that healed within 9 weeks, the same animal that had been shown to secrete low levels of IFN-gamma. The results demonstrate high cross-reactivity between L. donovani and L. major and that L. donovani protects against L. major infections. This finding is important for vaccine development studies against leishmaniasis.

Animals↗

Palisading neutrophilic granulomatous dermatitis in a Japanese patient with Wegener's granulomatosis.

Wegener's granulomatosis (WG) is an etiologically obscure entity with multiple systemic manifestations. Recently, cytoplasmic anti-neutrophil cytoplasmic antibody (C-ANCA) has become recognized as a valuable adjunct in the diagnosis of this disorder. WG typically involves the upper airway, lungs, and kidneys, but any other organ can be involved, including the skin. We encountered a unique case in which a 27-year-old Japanese man with WG presented with various typical cutaneous manifestations. Purpuric skin lesions and erythematous rash on the lower extremities progressively involved and changed into a necrotizing ulceration on his toe. Additionally, several nodules developed on the extensor surfaces of his elbows. His serum C-ANCA level increased remarkably. Leukocytoclastic vasculitis, the most common histopathological finding in WG patients, was detected in a purpuric lesion on his hand. A biopsy of a nodule on his elbow revealed palisading epithelioid histiocyte granulomas with features of leukocytoclastic vasculitis. The distinctive pattern of papules has been referred to as "palisading neutrophilic granulomatous dermatitis". An open lung biopsy confirmed WG with focal necrotizing granuloma. A renal biopsy demonstrated necrotizing vasculitis and crescentic glomerulonephritis. He showed a good response to oral corticosteroids and cyclophospamide with total remission of symptoms. We believe that a careful balance between the clinical manifestations and the histopathological evidence allows for timely treatment of WG, which may prevent serious morbidity or death. Although uncommon, WG can present with various types of accompanying cutaneous lesions. Therefore, clinicians should keep this diagnosis in mind when presented with these manifestations.

Adult↗

A retrospective review of the management of Dupuytren's nodules.

PURPOSE: To evaluate the progression of Depuytren's nodules with more than 6 years of follow-up study. METHODS: Fifty-nine patients who presented initially with Dupuytren's nodules returned for physical examination at an average follow-up period of 8.7 years (range, 6-15 y). Patients were questioned regarding family history of Dupuytren's disease, family ethnicity, alcohol consumption, smoking, liver disease, seizures, diabetes, and signs of systemic disease such as knuckle pads and plantar nodules. Physical examination evaluated disease state, loss of extension of the finger joints, and disease location. RESULTS: Thirty of the 59 patients with previously diagnosed isolated nodules developed a cord. Twenty-two percent of patients presented with bilateral disease and another 26% developed bilateral disease. Of those patients whose disease progressed 43% had European heritage, 37% had disease onset before the age of 50 years, 30% had bilateral disease, 23% had a family history of Dupuytren's disease, and 13% had plantar nodules. Five patients lost extension averaging 60 degrees at the metacarpophalangeal joint and 40 degrees at the proximal interphalangeal joint. Three of these 5 had surgical excision because they had a flexion contracture of the metacarpophalangeal or proximal interphalangeal joints averaging 60 degrees and 43 degrees , respectively. Another 7 patients did not meet standard criteria but had surgery for persistent pain associated with grasping objects (without contracture). All surgically treated patients had at least 1 risk factor and 7 patients had more than 1 risk factor. In 7 patients the Dupuytren's nodule had resolved at the time of follow-up evaluation. CONCLUSIONS: The progression of the nodular form of Dupuytren's disease to cord-like disease is common but not inevitable. This evaluation of Dupuytren's nodules has shown that at an average of 8.7 years after diagnosis 5 patients met standard surgical criteria of metacarpophalangeal contracture of greater than 30 degrees or any proximal interphalangeal contracture. Age of onset (before 50 years) is correlated most closely with disease progression, and the disease regressed in 7 patients (12%).

Adolescent↗

Juvenile temporal arteritis revisited.

We describe a case of arteritis involving the superficial temporal artery in an 8-year-old boy. After a 2-week prodrome of headache in the right temporal region, a painful pulsatile 6-mm nodule developed. No history of trauma or systemic disease was noted. The differential diagnosis included vasculitis or thrombosis of a vascular malformation of the temporal artery. The lesion was surgically excised for both diagnostic and cosmetic reasons. Histologic features of the nodule were diagnostic of juvenile temporal arteritis and characterized by non-giant cell granulomatous inflammation of the temporal artery, occlusive fibrous intimal proliferation, and microaneurysmal disruption of the media. At 12-month follow-up, the patient was well; no recurrent lesions or systemic disease was noted. Although rare, this disease should be recognized as arteritis that affects the external carotid circulation and should not be confused with classic giant cell temporal arteritis. If physicians are aware of this benign inflammatory disease of the temporal artery in children and young adults, unnecessary treatment will not be administered.

Child↗

Alfalfa NADH-dependent glutamate synthase: structure of the gene and importance in symbiotic N2 fixation.

Glutamate synthase (GOGAT), a key enzyme in ammonia (NH+4) assimilation, occurs as two forms in plants: a ferredoxin-dependent form (Fd-GOGAT) and an NADH-dependent form (NADH-GOGAT). These enzymes are encoded by distinct genes as evidenced by their cDNA and deduced amino acid sequences. This paper reports the isolation and characterization of a NADH-GOGAT gene from alfalfa (Medicago sativa L.), the first GOGAT gene to be isolated from a eukaryote. RNase protection and primer extension experiments map the transcription start site of NADH-GOGAT to nearly identical positions. The transcribed region of this gene, 12,214 bp, is comprised of 22 exons separated by 21 introns. The 2.7 kbp region 5' from the translation initiation site confers nodule-specific reporter gene activity when used in a chimeric beta-glucuronidase (GUS) construct and transformed into Lotus corniculatus and Medicago sativa. Both infected and uninfected cells display GUS activity. The abundance of NADH-GOGAT transcripts increases substantially in developing nodules of plants infected with effective rhizobia. However, this increase is not observed when nodules are induced by a variety of ineffective rhizobial strains. Thus, unlike many other plant genes involved in root nodule NH+4 assimilation, high levels of NADH-GOGAT expression are strictly associated with effective nodules indicating that NADH-GOGAT plays a central role in the functioning of effective root nodules. An alfalfa Fd-GOGAT PCR product showing greater than 85% identity to maize Fd-GOGAT was isolated and used to investigate the contribution of this enzyme to NH+4 assimilation in nodules. Fd-GOGAT mRNA was abundant in leaves and cotyledons but was not detected in alfalfa root nodules. Fd-GOGAT in alfalfa does not appear to play a significant role in symbiotic N2 fixation.

Amino Acid Sequence↗

Genetic analyses of Rhizobium meliloti exopolysaccharides.

We have recently obtained strong genetic evidence that the acidic Calcofluor-binding exopolysaccharide (EPS I) of Rhizobium meliloti Rm1021 is required for nodule invasion and possibly for later events in nodule development. Thirteen loci on the second megaplasmid have been identified that are required for, or affect, the synthesis of EPS I. Mutations in certain of these loci completely abolish the production of EPS I and result in mutants that form empty Fix- nodules. exoH mutants fail to succinylate their EPS I and form empty Fix- nodules. We have identified two unlinked regulatory loci, exoR and exoS, whose products play negative roles in the regulation of expression of the exo genes. We have recently discovered that R. meliloti has a latent capacity to synthesize a second exopolysaccharide (EPS II) that can substitute for the role(s) of EPS I in nodulation of alfalfa but not of other hosts. Possible roles for Rhizobium exopolysaccharides in nodulation are discussed.

Carbohydrate Sequence↗

Subcutaneous leiomyosarcoma developing in a radiation dermatitis.

A case of subcutaneous leiomyosarcoma developing in a skin affected by radiation dermatitis is presented. A 59-year-old patient had been suffering from radiation dermatitis induced by radiotherapy for a mediastinal tumor diagnosed at 24 years of age. Thirty-four years later, slow growing nodules developed in this lesion. Light and electron microscopic studies revealed that the tumor was a subcutaneous leiomyosarcoma. There is no previous report of a subcutaneous leiomyosarcoma developing in radiation dermatitis.

Dermatitis↗

Cavitary lung nodules associated with combination chemotherapy containing bleomycin.

We describe a patient in whom multiple cavitary lung nodules developed without infectious symptoms after treatment with bleomycin, cisplatin and vinblastine combination chemotherapy for metastatic testis cancer. Open lung biopsy demonstrated extensive granuloma formation without evidence of malignancy or infection. This report extends the spectrum of bleomycin pulmonary toxicity to include cavitary pulmonary nodules associated with granuloma formation.

Antineoplastic Combined Chemotherapy Protocols↗

Evaluation of risk in autonomously functioning thyroid nodules.

Based on iodine deficiency in Germany autonomous functioning nodules develop and are loaded with the risk of hyperthyroidism. Even subclinical hyperthyroidism has health implications including general symptoms, diseases of the cardiovascular system (atrial fibrillation, risk of thrombembolism, stroke and other embolic events) and a reduction of axial bone mineral density. Fortunately functional nodules have a very low probability of being malignant. However, there exist a coexistence of thyroid carcinoma outside the hot nodule.

Cardiovascular Diseases↗

Nodular desmoplastic variant of trichoblastoma.

A case of an unusual, previously unreported, nodular desmoplastic variant of trichoblastoma (hair germ neoplasm) in the scalp of a 65-year-old woman is reported. Four discrete nodules were present in close proximity at presentation. Another nodule developed in the same vicinity 3 1/2 years after excision, and there was no further recurrence 2 1/2 years later. The tumors were located in the dermis, made up of cords of primitive hair germ-like structures radiating down from discrete hyalinized nodules located in the superficial dermis. There were small foci of keratinization or mucoid change within the epithelial islands. Toward the deep portion of the tumor, the epithelial cells became spindly and grew in a more dispersed pattern, within a desmoplastic stroma. There was also infiltration of the epineurium of the cutaneous nerves and arector pili muscle. This uncommon benign neoplasm of hair germ merits wider recognition; in particular, this desmoplastic variant may mimic invasive carcinoma, morphea-type basal cell carcinoma, desmoplastic trichoepithelioma, and desmoplastic trichilemmoma. Distinguishing this variant from the former two possibilities is particularly important in order to avoid unduly aggressive treatment.

Aged↗

NADH-glutamate synthase in alfalfa root nodules. Genetic regulation and cellular expression.

NADH-dependent glutamate synthase (NADH-GOGAT; EC 1.4.1.14) is a key enzyme in primary nitrogen assimilation in alfalfa (Medicago sativa L.) root nodules. Here we report that in alfalfa, a single gene, probably with multiple alleles, encodes for NADH-GOGAT. In situ hybridizations were performed to assess the location of NADH-GOGAT transcript in alfalfa root nodules. In wild-type cv Saranac nodules the NADH-GOGAT gene is predominantly expressed in infected cells. Nodules devoid of bacteroids (empty) induced by Sinorhizobium meliloti 7154 had no NADH-GOGAT transcript detectable by in situ hybridization, suggesting that the presence of the bacteroid may be important for NADH-GOGAT expression. The pattern of expression of NADH-GOGAT shifted during root nodule development. Until d 9 after planting, all infected cells appeared to express NADH-GOGAT. By d 19, a gradient of expression from high in the early symbiotic zone to low in the late symbiotic zone was observed. In 33-d-old nodules expression was seen in only a few cell layers in the early symbiotic zone. This pattern of expression was also observed for the nifH transcript but not for leghemoglobin. The promoter of NADH-GOGAT was evaluated in transgenic alfalfa plants carrying chimeric beta-glucuronidase promoter fusions. The results suggest that there are at least four regulatory elements. The region responsible for expression in the infected cell zone contains an 88-bp direct repeat.

Amino Acid Oxidoreductases↗

Morphogenesis of septa in hepatic fibrosis induced by choline deficiency in rats. Correlation between ito cells, pericellular fibrosis, and septa.

Examination of hepatic fibrosis induced by choline deficient diet showed the histogenesis of early fibrosis and septa formation. Male rats were fed a choline deficient diet. Groups of three were sacrificed after 1, 2, 3, 6, 8, 12, and 13 weeks. Control groups of three rats were sacrificed after 6, 8, and 12 weeks. All choline deficient rats showed initial steatosis around the portal tracts. The fat droplets were small at first and gave a microvesicular appearance. As they grew, they gathered around the terminal hepatic venule by the 3rd week. Then, reticular fibers developed around the hepatocytes in the centrolobular area and fibroses connected the terminal hepatic venules. The fat droplets became larger and made nodular masses around the portal tract. These compressed the fibroses to the periphery where they formed septa. After 12 weeks, regenerating nodules developed. Electron microscopy showed that fat storing cells helped cause pericellular fibrosis.

Animals↗

Studies on the antitumor activities of pyrimidinone-interferon inducers. I. Effect against artificial and spontaneous lung metastases of murine tumors.

Since pyrimidinone compounds induce interferon production in several animal species and have potent antivirus activities, it appeared important to determine whether these compounds could also induce antitumor activities in their recipients. Pyrimidinone compounds 2-amino-5-bromo-6-methyl-4-pyrimidinone (ABMP), 2-amino-5-brome-6-phenyl-4-pyrimidinone (ABPP), and 2-amino-5-iodo-6-phenyl-4-pyrimidinone (AIPP) were studied for their activities against artificial lung metastases of the weakly immunogenic spontaneous fibrosarcoma NFSa, the moderately immunogenic spontaneous mammary carcinoma MCa-K, and the strongly immunogenic 3-methylcholanthrene-induced fibrosarcoma FSa syngeneic to inbred C3Hf/Kam mice. In addition, the therapeutic efficacy of ABPP and AIPP was also determined against spontaneous lung metastases of NFSa. ABPP and AIPP given ip at 250 mg/kg for 2 or 3 consecutive days before or after iv inoculatin of NFSa, FSa, or MCa-K cells greatly reduced the number of tumor nodules developed in the lungs. ABMP, however, was considerably less effective. ABPP and AIPP were also effective in therapy of spontaneous lung metastases of NFSa, especially when these compounds were given before surgical removal of the primary tumor. Neither ABPP nor AIPP was effective against tumor nodules growing in whole-body irradiated (WBI) mice, but both protected mice against enhancement of lung metastasis formation induced by exposure to whole-body irradiation. ABPP was more effective than AIPP in inducing production of interferon in normal mice. When treated with ABPP, WBI mice, however, were unable to produce interferon. These results show that 6-phenyl-pyrimidinone compounds induce strong antitumor activities in mice, which correlated with neither tumor immunogenicity nor the ability of these agents to induce interferon, but which depended on the immune status of the tumor host.

Animals↗

Lupus erythematosus panniculitis in a patient with autoimmune hepatitis.

Lupus erythematosus panniculitis (profundus), a rare variant of chronic panniculitis, sometimes develops during the course of discoid lupus erythematosus or systemic lupus erythematosus. A 61-year-old woman had suffered from autoimmune hepatitis type I for 5 years. Prednisolone had been administered as maintenance therapy and her hepatitis had been well controlled. However, asymptomatic erythematous indurated nodules developed symmetrically in both pre-auricular regions, and skin biopsy revealed lupus erythematosus panniculitis (profundus). Increase in dosage of prednisolone resolved the skin lesion, leaving depressed atrophic scars. This is the first report of lupus erythematosus panniculitis complicating autoimmune hepatitis.

Biopsy↗

Correlation between circulating cancer cells and incidence of metastases.

Quantitative aspects of the behaviour of B16 melanoma and Lewis lung carcinoma cells during the post-intravasation stages of metastasis were examined in relation to their spontaneous metastatic potential. Cancer cells were isolated from the blood of mice bearing i.m. tumours throughout tumour growth using a novel discontinuous gradient centrifugation technique. Four times more Lewis carcinoma cells than B16 melanoma cells were shed into the circulation, although the numbers of cells shed from either tumour were orders of magnitude more than the numbers of spontaneous pulmonary metastases which developed. Greater numbers of Lewis carcinoma cells were also shed as clumps and with leukocytes attached. However, although similar numbers of radiolabelled melanoma and Lewis carcinoma cells were arrested in the lungs after i.v. injection, fewer carcinoma cells were retained there and 12 times fewer Lewis carcinoma nodules developed in the lungs following injection of non-radiolabelled cells. It appears that the low lung colonization potential of the Lewis lung carcinoma is compensated for during spontaneous metastases by the numbers of cells shed from primary tumours as single cells and as clumps.

Animals↗

Structural characterization of a K-antigen capsular polysaccharide essential for normal symbiotic infection in Rhizobium sp. NGR234: deletion of the rkpMNO locus prevents synthesis of 5,7-diacetamido-3,5,7,9-tetradeoxy-non-2-ulosonic acid.

Many early molecular events in symbiotic infection have been documented, although factors enabling Rhizobium to progress within the plant-derived infection thread and ultimately survive within the intracellular symbiosome compartment as mature nitrogen-fixing bacteroids are poorly understood. Rhizobial surface polysaccharides (SPS), including the capsular polysaccharides (K-antigens), exist in close proximity to plant-derived membranes throughout the infection process. SPSs are essential for bacterial survival, adaptation, and as potential determinants of nodulation and/or host specificity. Relatively few studies have examined the role of K-antigens in these events. However, we constructed a mutant that lacks genes essential for the production of the K-antigen strain-specific sugar precursor, pseudaminic acid, in the broad host range Rhizobium sp. NGR234. The complete structure of the K-antigen of strain NGR234 was established, and it consists of disaccharide repeating units of glucuronic and pseudaminic acid having the structure -->4)-beta-d-glucuronic acid-(1-->4)-beta-5,7-diacetamido-3,5,7,9-tetradeoxy-l-glycero-l-manno-nonulosonic acid-(2-->. Deletion of three genes located in the rkp-3 gene cluster, rkpM, rkpN, and part of rkpO, abolished pseudaminic acid synthesis, yielding a mutant in which the strain-specific K-antigen was totally absent: other surface glycoconjugates, including the lipopolysaccharides, exopolysaccharides, and flagellin glycoprotein appeared unaffected. The NGRDeltarkpMNO mutant was symbiotically defective, showing reduced nodulation efficiency on several legumes. K-antigen production was found to decline after rhizobia were exposed to plant flavonoids, and the decrease coincided with induction of a symbiotically active (bacteroid-specific) rhamnan-LPS, suggesting an exchange of SPS occurs during bacterial differentiation in the developing nodule.

Antigens, Bacterial↗

[Hemithyroidectomy in a unilateral goiter: a valid therapeutic option. Review of the contralateral hemithyroid ten years after treatment].

INTRODUCTION: Nodular goiter (NG) is frequent among the general population and is considered a diffuse disease. Although NGs are rarely unilateral, they pose a dilemma in terms of the extent of the thyroidectomy. The aim of the present study was to evaluate the remaining thyroid in patients with NG compared with those with follicular adenoma who underwent hemithyroidectomy. PATIENTS AND METHODS: Patients who underwent surgery for unilateral NG with over 10 years of postoperative follow-up and normal findings on ultrasonography of the contralateral thyroid lobe were selected to form the study group (SG). Patients with follicular adenoma (with normal contralateral ultrasonography) who underwent hemithyroidectomy during the same period were selected to form the control group (CG). The selected patients underwent clinical, laboratory and ultrasound examinations. Both groups were compared statistically. No significant differences were found in age, gender, anesthetic risk, side, postoperative complications, length of hospital stay, or postoperative outcome. RESULTS: Less than 10% of the patients reported symptoms, and all symptoms were of little significance. Ultrasonographic nodules were found in the remaining thyroid lobe in 70% of patients in the SG and in 60% of those in the CG, with no statistically significant differences. The mean size of the largest nodule was 13.58 +/- 8.01 in the SG and 9.15 +/- 5.93 in the GC (p = 0.048). No differences were found in the anterior-posterior, transverse or longitudinal diameters of the remaining lobe. None of the patients underwent reintervention for nodular disease. CONCLUSIONS: After hemithyroidectomy, both groups of patients developed nodules in the remaining thyroid lobe, with no statistically significant differences. Hemithyroidectomy due to unilateral NG involves less risk to the patient and therefore we consider it to be a valid option. Long-term ultrasonographic follow-up seems advisable.

Adult↗