Molecular genetic methods in the diagnosis of hematologic neoplasms

Leukemias and lymphomas are monoclonal neoplasms that arise as a result of molecular abnormalities. These abnormalites are diverse but can be grouped into two general categories, chromosomal translocations that usually result in oncogene activation and inactivation of tumor suppressor genes. Recent advances in our understanding of chromosomal translocations have led to improved classification of leukemias and lymphomas. For example, the t(9;22)(q34;q11) is now considered a defining feature of chronic myeloid leukemia, and the t(2;5)(p23;q35) defines a clinically and biologically unique subset of anaplastic large cell lymphomas. In this review, we focus on chromosomal translocations in hematologic neoplasms and the techniques used for their detection. We also briefly discuss tumor suppressor genes and assessment of clonality in lymphoid neoplasms.     

Localization of 5-hydroxytryptamine-like immunoreactive cells and nerve fibers in the rat female reproductive system.

The presence of 5-hydroxytryptamine (5-HT)-like immunoreactivity (IR) was studied in the rat female reproductive system using polyclonal antibodies directed against 5-HT. Moreover, 5-HT levels in the ovary, oviduct, uterus, and cervix were measured by high-pressure liquid chromatography with electrochemical detection. The highest 5-HT concentrations were found in the oviduct, followed in descending order by the cervix, the ovary, and the uterus. Most 5-HT-like IR was observed in the cytoplasm of mast cells. These cells were found in the connective tissue around the fimbria, in the oviduct, in the uterus, and in the ovary. Mast cells are clustered in the proximity of the parenchymal blood vessels. Moreover, a few 5-HT-like nerve fibers were found distributed mainly perivascularily in the uterine cervix and in the uterine horns as well as in the oviduct. IR nerve fibers were rarely seen within the ovary. The present data provide direct evidence that 5-HT in the female reproductive system not only is associated with mast cells but is located in nerve fibre-like structures as well. The functional significance of this probable 5-HT-ergic innervation of the female reproductive tract discovered in the present study should be clarified in future investigations.     

Two new mutations and three novel polymorphisms in the<Emphasis Type="Italic"> RB1</Emphasis> gene in Ecuadorian patients

RB1 is the gene responsible for retinoblastoma, the most common malignant intraocular tumor of infancy and early childhood. There are no reports about this gene in Ecuadorian populations, and only a few studies have been published in Latin America about this subject. There is a spectrum of more than 370 mutations described in the RB1 gene mutation database (http://www.d-lohmann.de/Rb/mutations.html), and alterations have been found in 25 of the 27 exons. During the exon-by-exon analysis of 31 tumor and blood samples from Ecuadorian patients, we found two new mutations and three novel polymorphisms. One of the polymorphisms is located in intron 26 where no alterations of the gene have been described previously. The polymorphisms were found in all of the patients tumor samples, but not in normal population, suggesting there might be a relationship between these polymorphisms and the development of retinoblastoma in the Ecuadorian population.The nucleotide sequence data reported are available in the GenBank database under the accession numbers: AY243567, AY260472, AY260473, AY273783     

Evaluation of Urea-motility-indole medium for recognition and differentiation of Salmonella and Shigella species in stool cultures.

A semisolid urea-motility-indole medium designed for detection in Enterobacteriaceae of urease activity, motility, and indole production in one tube was prepared and evaluated. The formulation of the medium was similar to that of Christensen urea agar, but the agar concentration was 0.2%, and 1% tryptone was added. Results with 687 strains of Enterobacteriaceae were the same as those obtained with standard test media (98% overall agreement). The urea-motility-indole medium was also used in combination with Kligler iron agar for the recognition and differentiation of Salmonella and Shigella species from colonies picked from plating media in fecal cultures. This combination was compared with the combination of Kligler iron agar and lysine iron agar with 507 strains of non-lactose-fermenting Enterobacteriaceae. Although both combinations enabled the presumptive recognition and differentiation of Salmonella and Shigella species, an analysis of data indicated that the combination of Kligler iron agar and urea-motility-indole medium performed better than the combination of Kligler iron agar and lysine iron agar in detecting Salmonella and Shigella species.     

Initial treatment of the early stages (I, II) of supraglottic squamous cell carcinoma: partial laryngectomy versus radiotherapy

The purpose of this study was to define the treatment of choice (partial laryngectomy vs radiotherapy) in the early stage of supraglottic squamous cell cancer (ESSC). One hundred and fifteen patients with ESSC were treated with either partial laryngectomy (25 patients) or with radiotherapy(90 patients) between January 1984 and December 1996. All patients had a follow-up of over ¶29 months. Radiotherapy (RT) had a local control rate of 79%, which increased to 90% with salvage surgery, and a high larynx preservation rate (83%). Partial laryngectomy (PL) offered a better initial local control rate of 84%, which increased to 88% with salvage surgery, and functional results were also good (80%). No statistically significant differences were found between RT and PL. RT was less costly, showed better suitability for treatment, produced moderate morbidity and sequelae, and local recurrence was easier to rescue. However, it is a once-only application technique. PL showed higher immediate postoperative morbidity, higher cost and lower suitability for treatment but had fewer sequelae, offered the best initial local control and is multi-applicable. No clear oncological arguments were found in our series to define whether PL or RT is the treatment of choice for ESSC. Both are effective therapies. Secondary factors such as suitability for treatment, morbidity, cost and applicability should be individually evaluated when choosing the type of treatment. As the laser endoscopic approach decreases morbidity and costs and makes the condition more suitable for treatment, it could be the treatment of choice for ESSC, in cases where local tumoral extent and larynx exposure allow radical excision.     

Chromosome painting in plants: in situ hybridization with a DNA probe from a specific microdissected chromosome arm of common wheat.

We report here on the successful painting of a specific plant chromosome within its own genome. Isochromosomes for the long arm of chromosome 5 of the wheat B genome (5BL) were microdissected from first meiotic metaphase spreads of a monoisosomic 5BL line of the common wheat Triticum aestivum cv. Chinese Spring. The dissected isochromosomes were amplified by degenerate oligonucleotide-primed PCR in a single tube reaction. The amplified DNA was used as a complex probe mixture for fluorescent in situ hybridization on first meiotic metaphase spreads of lines carrying 5BL as a distinctive marker. Hybridization signals were observed, specifically, along the entire 5BL. In some of the cells, labeling was also detected in two bivalents, presumably those of the 5B "homoeologues" (partial homologues) found in common wheat (5A and 5D). The probe also revealed discrete domains in tapetal nuclei at interphase, further supporting the probe's high specificity. These data suggest that chromosome and homoeologous group-specific sequences are more abundant in 5BL than genome-specific sequences. Chromosome-painting probes, such as the one described here for 5BL, can facilitate the study of chromosome evolution in polyploid wheat.     

Who is Hispanic? Definitions and their consequences.

What is the appropriate method for classifying Spanish-speaking-origin inhabitants of the United States? This paper presents relevant data from the first wave of a longitudinal study of adolescents in the greater Miami area. As expected, the broadest definition--"up to third generation" Hispanic--identified the largest proportion of the sample as Hispanic, whereas parent self-report placed the smallest proportion into the Hispanic category. When policymakers are concerned about enumerating the entire Hispanic population, a definition broader than self-identification should be used; in estimating prevalence rates, however, the use of self-identification may be adequate.     

Clinical features of hepatic artery thrombosis after pediatric liver transplantation

Thrombosis of the hepatic artery (HAT) is a severe complication of liver transplantation, and most cases need regrafting. The aim of this study was to review our experience with this complication. From January 1986 through January 1992, 76 liver transplants were performed in 59 pediatric patients at the Children's Hospital La Paz, Madrid. The diagnosis of HAT was made in 12 cases (15.7%). The common patterns of clinical presentation were: fulminant liver necrosis (5), bile leak due to necrosis of the bile duct (4), and relapsing bacteremia (3). Clinical symptoms of fulminant liver necrosis started within the first 2 weeks after transplantation, with rapid deterioration and steep rises in SGOT and SGPT levels. All these patients were retransplanted on an urgent basis, but only 1 is alive 4 years later. Four patients developed bile leaks 13 to 60 days after transplantation; SGOT, SGPT, and total bilirubin were only slightly increased. Three children were retransplanted electively and are alive with a mean follow-up of 3 years. One exceptional patient had a Roux-en-Y jejunostomy and is doing well 30 months later with his original graft. The 3 remaining children had episodes of septicemia with hepatic abscess, liver infarction, and pleural effusion. Liver function tests were normal, with bilirubin levels below 2 mg/dl. All patients were retransplanted, but only 1 is alive and well 13 months later. In the present series, we found that early HAT produces fulminant clinical deterioration requiring an urgent regraft. Late HAT presenting with either infection or bile leak allows time for treatment by elective retransplantation. The best survival was obtained in the latter group. Correspondence to: J. Vázquez     

Health status and functional limitations in the elderly population of the University Region of Health of Puerto Rico: methodologic aspects and prevalence estimates

In order to plan the health services for the elderly population, it is necessary to quantify their health status and their functional capacity. In Puerto Rico, few epidemiological studies have been conducted regarding functional capacity and chronic diseases in the elderly population. One of the difficulties to undertake these studies is the high cost and risks to move this population for clinical exams, in addition to the methodology limitation of self-report in the elderly population. This study shows the use of logistic regression to estimate the prevalence of chronic diseases and functional capacity, when the observed data are not consisted with the planed sampling scheme. Four hundred and eighty-seven elderly persons (65 yr. and over) were interviewed in the municipalities of Canóvanas, Carolina, Loíza and Trujillo Alto in Puerto Rico, using a cross-sectional design. More than half of the elderly had visual problems (IC 95%: 54.8%, 63.8%), arthritis (IC 95%: 52.7%, 61.5%) and hypertension (IC 95%: 47.3%, 56.3%). In the case of hypertension, significant differences (p < 0.05) by sex were observed, where women reported a higher prevalence than men. One of the higher prevalences in the functional capacity status was with urine accidents. More than one-third of the population is estimated to have this problem (IC 95%: 34.7%, 43.4%). One-fourth of the population had limitations with going out for shopping (IC 95%: 23.9%, 31.9%) and using public and private transportation (IC 95%: 19.5%, 26.9%). We conclude that the applied methodology was consistent with the estimation presented in the literature and statistics from the Puerto Rico Health Department. However, it is necessary to continue assessing the design and analytical methodology, in order to undertake consistent and periodic evaluations of the elderly population.