[Low-cost reconstructive treatment after resection of distal radius giant cell tumor]. / Tratamiento reconstructivo de bajo costo tras resección de tumor de células gigantes de radio distal.

INTRODUCTION: giant cell tumor (GCT) is a benign intramedullary bone tumor that frequently arises at the ends of long bones. After the distal femur and proximal tibia, the distal radius is the third most affected site with particularly aggressive tumors. Our objective is the presentation of the clinical case of a patient diagnosed with distal radius GCT classified in grade III of Campanacci who received a treatment adjusted to her economic possibilities. CASE REPORT: a 47-year-old female, without economic solvency and with some medical service. Treatment included block resection, reconstruction with distal fibula autograft, and radiocarpal fusion with blocked compression plate. Eighteen months later, the patient had good grip strength (80% on the healthy side) and had fine motor function in the hand. The wrist presented stability with pronation of 85o, supination of 80o, flexion-extension of 0o and a score of 6.7 in the DASH functional outcomes assessment questionnaire. His radiological evaluation five years after his surgery continued with no evidence of local recurrence and pulmonary involvement. CONCLUSION: the result in this patient, together with the published data, indicate that the block tumor resection technique, plus distal fibula autograft and arthrodesis with blocked compression plate provide an optimal result of functionality for the grade III distal radial tumor at low cost.

INTRODUCCIÓN: el tumor de células gigantes (TCG) es un tumor óseo intramedular benigno que surge con frecuencia en los extremos de los huesos largos. Después del fémur distal y la tibia proximal, el radio distal es el tercer sitio más afectado con tumores, particularmente agresivos. Nuestro objetivo es la presentación del caso clínico de una paciente con diagnóstico de TCG de radio distal clasificada en grado III de Campanacci que recibió un tratamiento ajustado a sus posibilidades económicas. REPORTE DE CASO: paciente femenino de 47 años, sin solvencia económica y sin ningún servicio médico. El tratamiento incluyó resección en bloque, reconstrucción con autoinjerto de peroné distal y artrodesis radiocarpiana con placa de compresión bloqueada. Dieciocho meses después, la paciente presentaba una buena fuerza de prensión (80% respecto al lado sano) y tenía una función motora fina en la mano. La muñeca presentó estabilidad con pronación de 85o, supinación de 80o, flexión-extensión de 0o y una puntuación de 6.7 en el cuestionario de evaluación de resultados funcionales DASH. Su evolución radiológica a cinco años después de su cirugía continuó sin datos de recidiva local y afectación pulmonar. CONCLUSIÓN: el resultado en esta paciente, junto con los datos publicados, indican que la técnica de resección tumoral en bloque, más el autoinjerto de peroné distal y la artrodesis con placa de compresión bloqueada proporcionan un resultado óptimo de funcionalidad para el tumor radial distal grado III a bajo costo.

Multicenter longitudinal study of B-lymphocyte depletion in refractory systemic lupus erythematosus: the LESIMAB study.

OBJECTIVE: This study aimed to investigate the effectiveness and safety of single and repeated courses of rituximab in patients with refractory lupus. METHODS: LESIMAB is a multicenter, retrospective, longitudinal study of lupus patients who have not responded to standard therapy and have been treated with rituximab. Response rates at six months and at follow-up were defined as efficacy outcomes. Complete response was defined as a SELENA-SLEDAI score ≤ two and a SELENA-SLEDAI Flare Index of zero. Partial response was defined as a reduction in the SELENA-SLEDAI score of ≥four points with no new or worsening of symptoms. Adverse events were collected. RESULTS: Seventy-three (62.9%) of 116 patients achieved a response at six months (complete in 22 and partial in 51). Ninety-seven (77.6%) of 128 patients achieved a response after a mean follow-up of 20.0 ± 15.2 months (complete in 50 and partial in 47). High baseline SLEDAI score, previous treatment with ≥100 mg/day prednisone, and no history of severe hematologic flare were associated with response after the first treatment course. The median time to response was 6.5 months (95% CI, 5.0-8.0). Thirty-seven patients (38.1%) relapsed after the first infusion. The flare was severe in seven cases and mild to moderate in 29 cases. Serious infection rate was 12.6/100 patient-years. A schedule of four weekly doses was associated with more serious infections. Six patients died: two of infection and four of lupus complications. CONCLUSION: Rituximab can be an effective treatment option for patients who have refractory lupus with severe or life-threatening disease with an acceptable tolerance profile.

[Applications of monoclonal antibodies and biotechnology products in the treatment of chronic inflammatory diseases]. / Aplicaciones de anticuerpos monoclonales y productos de biotecnología en el tratamiento de las enfermedades inflamatorias crónicas.

In recent years we have noticed the arrival of biological drugs for the treatment of rheumatoid arthritis (RA), Crohn's disease (CD), psoriasis, and other chronic inflammatory diseases. Those drugs are produced with biotechnology methods and are defined as biologicals because of they work on the immune system. Different cellular groups and inflammation mediators participate in the inflammatory process, all of them susceptible of a therapeutic approach; they are so-called biological targets. Inhibition of TNF and interleukina 1 (IL-1) has proven effective for the control of inflammation in diseases as RA or CD. At present we have two types of inhibitors of TNF, specific monoclonal antibodies (infliximab, adalimumab) and cellular receptors (etanercept) and an IL-1 inhibitor (anakinra). The use of TNF inhibitors has given rise to a substantial change in the treatment of RA and CD because of its effectiveness. Together with this beneficial effect, an increase of infections (some of them severe) has occurred, especially tuberculosis. Other side effects that can be considered infrequent include demyelinization, heart failure, blood dyscrasias and lymphomas, which means that a thorough knowledge of these drugs is necessary for their use. Other potential biological drugs still in investigational phase are mentioned.

Aplicaciones de anticuerpos monoclonales y productos de biotecnología en el tratamiento de las enfermedades inflamatorias crónicas / Applications of monoclonal antibodies and biotechnology products in the treatment of chronic inflammatory diseases

En los últimos años hemos asistido a la llegada de fármacos biológicos para el tratamiento de artritis reumatoide, enfermedad de Crohn, psoriasis y otras enfermedades inflamatorias crónicas. Se definen como biológicos aquellos fármacos obtenidos por métodos de biotecnología y que actúan sobre el sistema inmunitario. En el proceso inflamatorio intervienen diferentes grupos celulares y mediadores de inflamación susceptibles de actuación terapéutica, son las llamadas dianas biológicas. La inhibición del factor de necrosis tumoral (TNF) e interleucina 1 se ha probado eficaz en el control de la inflamación en enfermedades como artritis reumatoide o enfermedad de Crohn. En la actualidad disponemos de dos tipos de inhibidores del TNF, anticuerpos monoclonales específicos (infliximab, adalimumab) y receptores celulares (etanercept) y un inhibidor de interleucina 1 (anakinra). El uso de inhibidores de TNF ha significado un cambio sustancial en el tratamiento de artritis reumatoide y enfermedad de Crohn por su eficacia. Junto a este efecto beneficioso se ha producido un aumento de infecciones, algunas de curso grave, especialmente tuberculosis. Otros efectos secundarios que se pueden considerar infrecuentes incluyen desmielinización, insuficiencia cardíaca, discrasias sanguíneas y aparición de linfomas, por lo que es necesario un profundo conocimiento de estos fármacos para su manejo. Se mencionan otros potenciales fármacos biológicos en fase de estudio

In recent years we have noticed the arrival of biological drugs for the treatment of rheumatoid arthritis (RA), Crohn’s disease (CD), psoriasis, and other chronic inflammatory diseases. Those drugs are produced with biotechnology methods and are defined as biologicals because of they work on the immune system. Different cellular groups and inflammation mediators participate in the inflammatory process, all of them susceptible of a therapeutic approach; they are so-called biological targets. Inhibition of TNF and interleukina 1 (IL-1) has proven effective for the control of inflammation in diseases as RA or CD. At present we have two types of inhibitors of TNF, specific monoclonal antibodies (infliximab, adalimumab) and cellular receptors (etanercept) and an IL-1 inhibitor (anakinra). The use of TNF inhibitors has given rise to a substantial change in the treatment of RA and CD because of its effectiveness. Together with this beneficial effect, an increase of infections (some of them severe) has occurred, especially tuberculosis. Other side effects that can be considered infrequent include demyelinization, herat failure, blood dyscrasias and lymphomas, which means that a thorough knowledge of these drugs is necessary for their use. Other potential biological drugs still in investigational phase are mentioned

Blakeslea trispora genes for carotene biosynthesis.

We cloned the carB and carRA genes involved in beta-carotene biosynthesis from overproducing and wild-type strains of Blakeslea trispora. The carB gene has a length of 1,955 bp, including two introns of 141 and 68 bp, and encodes a protein of 66.4 kDa with phytoene dehydrogenase activity. The carRA gene contains 1,894 bp, with a single intron of 70 bp, and encodes a protein of 69.6 kDa with separate domains for lycopene cyclase and phytoene synthase. The estimated transcript sizes for carB and carRA were 1.8 and 1.9 kb, respectively. CarB from the beta-carotene-overproducing strain B. trispora F-744 had an S528R mutation and a TAG instead of a TAA stop codon. The overproducing strain also had a P143S mutation in CarRA. Both B. trispora genes could complement mutations in orthologous genes in Mucor circinelloides and could be used to construct transformed strains of M. circinelloides that produced higher levels of beta-carotene than did the nontransformed parent. The results show that these genes are conserved across the zygomycetes and that the B. trispora carB and carRA genes are functional and potentially useable to increase carotenoid production.

Biotechnological lycopene production by mated fermentation of Blakeslea trispora.

A semi-industrial process (800-l fermentor) for lycopene production by mated fermentation of Blakeslea trispora plus (+) and minus (-) strains has been developed. The culture medium was designed at the flask scale, using a program based on a genetic algorithm; and a fermentation process by means of this medium was developed. Fermentation involves separate vegetative phases for (+) and (-) strains and inoculation of the production medium with a mix of both together. Feeding with imidazole or pyridine, molecules known to inhibit lycopene cyclase enzymatic activity, enhanced lycopene accumulation. Different raw materials and physical parameters, including dissolved oxygen, stirring speed, air flow rate, temperature, and pH, were checked in the fermentor to get maximum lycopene production. Typical data for the fermentation process are presented and discussed. This technology can be easily scaled-up to an industrial application for the production of this carotenoid nowadays widely in demand.

Structural and phylogenetic analysis of the gamma-actin encoding gene from the penicillin-producing fungus Penicillium chrysogenum.

The nucleotide sequence of a 2994-bp genomic fragment, including the gamma-actin encoding gene from Penicillium chrysogenum, has been determined, showing an open reading frame (ORF) of 1756 bp interrupted by five introns with fungal consensus splice-site junctions. The 5' untranslated region contains a consensus TATA box, five CAAT motifs, and two large pyrimidine stretches. The predicted protein (375 amino acids) revealed high identity to gamma-actins from fungi (>90%), and gene phylogenies support the grouping of P. chrysogenum actin close to those from the majority of the filamentous fungi. The actA gene is present as a single copy in the genome of P. chrysogenum, and its expression is constitutive during penicillin fermentation, showing a single 1.4-kb transcript.

[Medicine and history. Concerning of the case of a man with a tumour on his forehead]. / Medicina e historia. A propósito del caso de un hombre con un tumor en la frente.

This case report is about the sickness of a 59 year old priest, politician and war veteran who died in 1844. The case history is based on the detailed report of his doctor, gathered in a diary and later published. The collected clinical data relied exclusively on touch and observation. The patient's illness started with a painful lump on his forehead that was in part excised. Surgical exploration revealed a soft tissue mass that bled easily and involved the frontal bone. In the following months, the lesion, which had been treated with complex topical medications, became ulcerated and extended to the orbit and the chin. The patient died postrated and in severe undernutrition one year after the onset of the symptoms. The case discussion, presented in the format of a clinicopathological conference, concluded that a metastasis of a renal cell carcinoma or an osseous lymphoma were the more likely diagnoses.

Medicina e historia. A propósito del caso de un hombre con un tumor en la frente. / [Medicine and history. Concerning of the case of a man with a tumour on his forehead]

This case report is about the sickness of a 59 year old priest, politician and war veteran who died in 1844. The case history is based on the detailed report of his doctor, gathered in a diary and later published. The collected clinical data relied exclusively on touch and observation. The patients illness started with a painful lump on his forehead that was in part excised. Surgical exploration revealed a soft tissue mass that bled easily and involved the frontal bone. In the following months, the lesion, which had been treated with complex topical medications, became ulcerated and extended to the orbit and the chin. The patient died postrated and in severe undernutrition one year after the onset of the symptoms. The case discussion, presented in the format of a clinicopathological conference, concluded that a metastasis of a renal cell carcinoma or an osseous lymphoma were the more likely diagnoses.

Overexpression of the lat gene in Nocardia lactamdurans from strong heterologous promoters results in very high levels of lysine-6-aminotransferase and up to two-fold increase in cephamycin C production.

The level of lysine-6-aminotransferase (encoded by the lat gene), an enzyme that commits lysine to the cephamycin biosynthesis pathway, is very low in wild type Nocardia lactamdurans. Two lat overexpression systems (pAMEXlat and pSAFlat) were constructed to express the promoterless lat gene of N. lactamdurans from the strong promoters amyP (of the alpha-amylase gene) and safP (of the secretion activating factor gene) of Streptomyces griseus. Both constructions led to very high levels of lysine-6-aminotransferase (between 8- and 15-fold) in the cells. Expression of lat from the amy promoter was optimal in glycerol-containing medium and was negatively regulated by glucose. The high levels of lysine-6-aminotransferase resulted in a 50-200% increase in cephamycin C production in the standard fermentation conditions. Onset of cephamycin C biosynthesis occurred at the same time in control and in lat-overexpressing strains, but the cephamycin production rate was clearly higher in transformants overexpressing the lat gene. Furthermore, HPLC analysis of cephamycin C in the culture broths revealed an early depletion of biosynthetic intermediates and an accumulation of cephamycin C when the lat gene was overexpressed. These results indicate that lysine-6-aminotransferase activity is limiting for cephamycin C biosynthesis under some culture conditions.

The gene encoding gamma-actin from the cephalosporin producer Acremonium chrysogenum.

The nucleotide sequence of a 3240-bp genomic fragment including the gamma-actin-encoding gene from Acremonium chrysogenum has been determined, showing an open reading frame of 1691 bp, interrupted by five introns with fungal consensus splice-site junctions. The untranslated regions of the actA gene contain a consensus TATA box, a CCAAT motif, pyrimidine stretches and the polyadenylation sequence AATAA. The predicted protein (375 amino acids) revealed high identity to gamma-actins from fungi (> 90%). Gene phylogenies constructed using DNA and protein sequences support the grouping of A. chrysogenum actin close to those from the majority of the filamentous fungi. The actA gene is present as a single copy in the genome of A. chrysogenum; and its expression level, opposite to pcbC and cefEF cephalosporin biosynthetic genes, was steady during cephalosporin fermentation, showing a single 1.4-kb transcript.

Inducing effect of diamines on transcription of the cephamycin C genes from the lat and pcbAB promoters in Nocardia lactamdurans.

The diamines putrescine, cadaverine, and diaminopropane stimulate cephamycin biosynthesis in Nocardia lactamdurans, in shake flasks and fermentors, without altering cell growth. Intracellular levels of the P7 protein (a component of the methoxylation system involved in cephamycin biosynthesis) were increased by diaminopropane, as shown by immunoblotting studies. Lysine-6-aminotransferase and piperideine-6-carboxylate dehydrogenase activities involved in biosynthesis of the alpha-aminoadipic acid precursor were also greatly stimulated. The diamine stimulatory effect is exerted at the transcriptional level, as shown by low-resolution S1 protection studies. The transcript corresponding to the pcbAB gene and to a lesser extent also the lat transcript were significantly increased in diaminopropane-supplemented cultures, whereas transcription from the cefD promoter was not affected. Coupling of the lat and pcbAB promoters to the reporter xylE gene showed that expression from the lat and pcbAB promoters was increased by addition of diaminopropane in Streptomyces lividans. Intracellular accumulation of diamines in Nocardia may be a signal to trigger antibiotic production.

Delta-1-piperideine-6-carboxylate dehydrogenase, a new enzyme that forms alpha-aminoadipate in Streptomyces clavuligerus and other cephamycin C-producing actinomycetes.

Delta-1-Piperideine-6-carboxylate (P6C) dehydrogenase activity, which catalyses the conversion of P6C into alpha-aminoadipic acid, has been studied in the cephamycin C producer Streptomyces clavuligerus by both spectrophotometric and radiometric assays. The enzyme has been purified 124-fold to electrophoretic homogeneity with a 26% yield. The native protein is a monomer of 56.2 kDa that efficiently uses P6C (apparent Km 14 microM) and NAD+ (apparent Km 115 microM), but not NADP+ or other electron acceptors, as substrates. The enzyme activity was inhibited (by 66%) by its end product NADH at 0.1 mM concentration. It did not show activity towards pyrroline-5-carboxylate and was separated by Blue-Sepharose chromatography from pyrroline-5-carboxylate dehydrogenase, an enzyme involved in the catabolism of proline. P6C dehydrogenase reached maximal activity later than other early enzymes of the cephamycin pathway. The P6C dehydrogenase activity was decreased in ammonium (40 mM)-supplemented cultures, as was that of lysine 6 amino-transferase. P6C dehydrogenase activity was also found in other cephamycin C producers (Streptomyces cattleya and Nocardia lactamdurans) but no in actinomycetes that do no produce beta-lactams, suggesting that it is an enzyme specific for cephamycin biosynthesis, involved in the second stage of the two-step conversion of lysine to alpha-aminoadipic acid.

Amy as a reporter gene for promoter activity in Nocardia lactamdurans: comparison of promoters of the cephamycin cluster.

Promoter probe vectors containing the pA origin of replication and the Streptomyces griseus promoterless amy gene (encoding alpha-amylase) as reporter have been constructed to study transcription initiation regions in Nocardia lactamdurans. In some of the promoter probe vectors the phage fd terminator has been introduced to avoid readthrough expression from upstream sequences. By using these vectors, four different transcription initiation regions of the cephamycin gene cluster have been studied in N. lactamdurans. The bla gene encoding a beta-lactamase has a relatively strong promoter. Two other separate promoters corresponding to the lat and cefD genes (encoding, respectively, lysine-6-aminotransferase and isopenicillin N-epimerase) showed weak transcription initiation ability. These two promoters are arranged in a bidirectional transcription initiation region located in the center of the cephamycin gene cluster. The cmcH gene (encoding 3-hydroxymethylcephem carbamoyltransferase) upstream region did not contain a functional promoter, suggesting that cmcH is transcribed as a part of a polycistronic mRNA. The native amy promoter is used very efficiently in N. lactamdurans, resulting in secretion of high levels of extracellular alpha-amylase.

Cistoadenoma seroso microquisto del pancreas aspectos clinicopatologicos y quirurgicos / Microcystic serous cystadenoma of pancreas. Clinicopathological and surgical features

The microcystic serous cystadenoma of pancreas or glycogen "rich"cystadenoma is a rare entity. Whe studiet five case of this cystadenoma in adult patients ages 47-68 (58 was the mean), four of wich were women (80 percent). The clinical presentation was varied. There was a prevalence of expansive manifestations with epigastric pain in three patients, and extrahepatic bile duct obstruction in other two. A distal tumour was revealed by the diagnostic methodology used (ultrasound and TAC) in three patients, and cephalic tumour in two, with a mean size of 8.8 cm. in diameter. A distal pancreatectomy was performed in two patients, a cephalic pancreatoduodenectomy was performed in one in relation with the presence of extrahepatic bile duct carcinoma, and the other two were treated with a partial cephalic pancreatectomy (enucleation). The nosological diagnose was post-surgical in all case of study. a prognosis for every case was dependat of the associated pathology.

Cistoadenoma seroso microquisto del pancreas aspectos clinicopatologicos y quirurgicos / Microcystic serous cystadenoma of pancreas. Clinicopathological and surgical features

The microcystic serous cystadenoma of pancreas or glycogen "rich"cystadenoma is a rare entity. Whe studiet five case of this cystadenoma in adult patients ages 47-68 (58 was the mean), four of wich were women (80 percent). The clinical presentation was varied. There was a prevalence of expansive manifestations with epigastric pain in three patients, and extrahepatic bile duct obstruction in other two. A distal tumour was revealed by the diagnostic methodology used (ultrasound and TAC) in three patients, and cephalic tumour in two, with a mean size of 8.8 cm. in diameter. A distal pancreatectomy was performed in two patients, a cephalic pancreatoduodenectomy was performed in one in relation with the presence of extrahepatic bile duct carcinoma, and the other two were treated with a partial cephalic pancreatectomy (enucleation). The nosological diagnose was post-surgical in all case of study. a prognosis for every case was dependat of the associated pathology. (AU)

[Microcystic serous cystadenoma of pancreas. Clinicopathological and surgical features]. / Cistoadenoma seroso microquístico del páncreas. Aspectos clinicopatológicos y quirúrgicos.

The microcystic serous cystadenoma of pancreas or glycogen "rich" cystadenoma is a rare entity. We studied five case of this cystadenoma in adult patients ages 47-68 (58 was the mean), four of which were women (80%). The clinical presentation was varied. There was a prevalence of expansive manifestations with epigastric pain in three patients, and extrahepatic bile duct obstruction in other two. A distal tumour was revealed by the diagnostic methodology used (ultrasound and TAC) in three patients, and cephalic tumour in two, with a mean size of 8.8 cm. in diameter. A distal pancreatectomy was performed in two patients, a cephalic pancreatoduodenectomy was performed in one in relation with the presence of extrahepatic bile duct carcinoma, and the other two were treated with a partial cephalic pancreatectomy (enucleation). The nosological diagnose was post-surgical in all case of study. A prognosis for every case was dependent of the associated pathology.

Overexpression of the Nocardia lactamdurans alpha-aminoadipyl-cysteinyl-valine synthetase in Streptomyces lividans. The purified multienzyme uses cystathionine and 6-oxopiperidine 2-carboxylate as substrates for synthesis of the tripeptide.

Formation of the tripeptide delta-(L-alpha-aminoadipyl)-L-cysteinyl-D-valine (Aad-Cys-Val) is catalyzed by a multienzyme peptide synthetase encoded by the pcbAB gene in producers of beta-lactam antibiotics. The pcbAB gene of Nocardia lactamdurans was overexpressed in Streptomyces lividans giving a high Aad-Cys-Val synthetase activity. The synthetase was purified 2785-fold to near homogeneity showing a molecular mass of 430 kDa by SDS/PAGE. The protein was identified in the gels with antibodies to Aad-Cys-Val synthetase and by the formation of aminoacyl-synthetase thioester complex with [14C]valine. The purified synthetase used alpha-aminoadipic acid or its lactam 6-oxopiperidine 2-carboxylic acid but was unable to use piperideine 6-carboxylic acid or pipecolic acid as substrates to form Aad-Cys-Val. L-Cystathionine, (2-amino-2-carboxyethyl)-L-homocysteine, was used as substrate and formed Aad-Cys-Val with the same efficiency as L-cysteine. The product of the reaction eluted with authentic Aad-Cys-Val. The synthetase preparation was able to hydrolyze L-cystathionine by a pyridoxal-phosphate-independent mechanism which is not inhibited by propargylglycine, to form Aad-Cys-Val.

New type of hexameric ornithine carbamoyltransferase with arginase activity in the cephamycin producers Streptomyces clavuligerus and Nocardia lactamdurans.

The ornithine carbamoyltransferases (OTCases) from the beta-lactam-producing actinomycetes Streptomyces clavuligerus and Nocardia lactamdurans have been purified to near-homogeneity by delta-N-phosphonoacetylornithine-Sepharose 4B affinity chromatography. The S. clavuligerus and N. lactamdurans OTCases monomers had a molecular mass of 37 kDa. The native OTCases of S. clavuligerus, N. lactamdurans and Streptomyces coelicolor had molecular masses of 248, 251 and 247 kDa respectively, which correspond to a hexameric structure. The apparent K(m) values for ornithine and carbamoylphosphate of the S. clavuligerus enzyme were respectively 2.3 and 6.0 mM at pH 8.0. The enzyme showed a reverse activity on citrulline and used lysine and putrescine as substrates. The hexameric complex showed coupled arginase-OTCase activities and was able to convert arginine into citrulline in a carbamoylphosphate-dependent manner. The requirement for carbamoylphosphate might prevent the arginase-OTCase complex from carrying out a futile cycle of arginine biosynthesis and degradation.