Performing bone marrow harvest on an outpatient basis: a single center UK experience

Bone marrow harvest (BMH) has historically been performed on an inpatient basis with a minimum of overnight inpatient stays. We commenced a program of outpatient (day case) BMH in 1999, performing 54 day case BMHs over a 3-year period. Of the total of 54 cases, 51 were known patients with hematological malignancies and 3 were healthy normal donors. Seven were excluded from day case BMH. Five (10.6%) of 47 patients/donors who were accepted for day case BMH required overnight admission. Two developed hypotension requiring intravenous fluid resuscitation. Two had excessive vomiting and 1 a difficult and prolonged harvest and was admitted at the request of the anesthetist. None of the patients admitted required more than overnight admission and 42 (89.4%) were discharged the same evening. In conclusion, day case BMH is safe, cost-effective, and reduces the pressure on inpatient beds.     

Development of rapid,sensitive one-tube duplex RT-PCR assay for specific and differential diagnosis of Chikungunya and dengue

Chikungunya and dengue, two arboviral infections are common in South-East Asia and their early clinical manifestations are very similar hence it is important to discriminate between them as early as possible for better clinical management. The aim of this study was to design a rapid, sensitive and specific method for the differential diagnosis of these two viruses simultaneously. A rapid one-tube duplex RT-PCR assay was developed that requires 110 min including RNA extraction, RT-PCR and agarose gel electrophoresis by using a novel Taq polymerase with high processivity. This one-tube duplex RT-PCR system with primers designed from the conserved regions of the genome allowed discrimination between the two viral groups. Bioinformatics analysis of the DNA sequences from PCR amplified products confirmed that this method was very specific and accurate. The time required for this duplex RT-PCR was comparable to the standard IgM capture ELISA method. This novel approach would help to diagnose specifically and accurately these two closely related arboviruses and enable early detection from blood. This method could be applied in resource limited settings, for surveillance in endemic regions or for routine epidemiological screening.     

Morphometric and Histological Study of Breast Lesions with Special Reference to Proliferative Activity and Invasiveness

Breast carcinoma is the most common cause of carcinoma death in women. Sometimes, difficulty arises to differentiate between premalignant lesions and carcinoma by routine histopathology. Our study was done to establish the role of morphometry and immunohistochemistry to solve this problem. In this study, total 60 cases of different breast lesions were included and 10 controls were also included to compare the results with the normal findings. They were studied by hematoxylin and eosin-stained sections for morphometry and routine histological study; as well as by proliferative markers such as proliferating cell nuclear antigen and p53. Invasiveness was studied using immunohistochemical staining with 34 βE12 monoclonal antibody. Statistically significant differences were found in morphometric parameters and in expression of proliferative markers between most of them. Morphometry and immunohistochemistry help in the proper diagnosis of different breast lesions that lie in the gray zone on routine histopathology.     

Gene sequence tags from Plasmodium falciparum genomic DNA fragments prepared by the "genease" activity of mung bean nuclease.

A genes-first approach to genome sequencing is described which efficiently generates gene sequence tags from genomic DNA. Mung bean nuclease (EC 3.1.30.1) cleaves the genomic DNA of many organisms before and after genes and within some introns. Analysis of gene sequence tags prepared from mung bean nuclease-digested Plasmodium falciparum DNA demonstrates that this method has several advantages over the popular cDNA expressed sequence tag approach. To date, 673 sequence tags containing over 215 kb of sequence have been generated from 400 clones. Sixty clones (15%) have significant similarity to sequences in the protein and translated nucleic acid data bases. These represent 51 unique genes, of which only 5 encode previously known P. falciparum proteins. The identified proteins include those expressed in erythrocytic, exoerythrocytic, and gametocytic stages of the parasite. Thirty percent of clones identified appear to carry complete coding regions. The spacer DNA separating genes is rarely cloned. These gene sequence tags will form a useful data base from which to initiate projects to develop new therapeutics, vaccines, and strategies to control human malaria.     

Endothelin-mediated remodeling in aortas of diabetic rats

BACKGROUND: Smooth muscle cells proliferation and extracellular matrix (ECM) protein deposition are key features of diabetic macroangiopathy. In the present study, we have studied the role of endothelin(A) (ET(A)) receptor, the predominant receptor on smooth muscle cells, in diabetes-induced vascular hypertrophy and remodeling. METHODS: Streptozotocin-induced diabetic rats were administrated a selective ET(A) receptor antagonist, TBC3214, for 26 weeks. Following treatment, aortas were harvested and subjected to gene expression and morphometric analyses. We quantified fibronectin (FN) and plasminogen activator inhibitor-1 (PAI-1) expression as indicators of increased ECM protein synthesis. ET-1, ET-3, transforming growth factor-beta1 (TGF-beta1) and angiotensinogen mRNA levels were measured to elucidate genes involved in FN expression. We have investigated an embryonic splice variant of FN, oncofetal FN, and nonmuscle myosin heavy chain (SMemb) as vascular remodeling indicators. RESULTS: Our results show that diabetes leads to upregulation of FN, PAI-1, ET-1, ET-3, TGF-beta1 and angiotensinogen mRNA levels in association with increased medial thickness. Immunohistochemical analyses revealed concurrent protein level changes. Diabetes also upregulated oncofetal FN and SMemb mRNA levels. Treatment with TBC3214 attenuated the mRNA levels of several genes and prevented increased medial thickness. CONCLUSIONS: These results indicate that diabetes-induced vascular hypertrophy and remodeling is associated with reexpression of embryonic forms of FN and myosin heavy chain. Such changes are ET-dependent and may be mediated via TGF-beta1 and angiotensin.