ABSTRACT
Knowledge of the exact cell content of frozen tissue samples is of growing importance in genomic research. We developed a microaliquoting technique to measure and optimize the cell composition of frozen tumor specimens for molecular studies. Frozen samples of 31 mesothelioma cases were cut in alternating thin and thick sections. Thin sections were stained and evaluated visually. Thick sections, i.e., microaliquots, were annotated using bordering stained sections. A range of cellular heterogeneity was observed among and within samples. Precise annotation of samples was obtained by integration and compared to conventional single face and "front and back"' section estimates of cell content. Front and back estimates were more highly correlated with block annotation by microaliquoting than were single face estimates. Both methods yielded discrepant estimates, however, and for some studies may not adequately account for the heterogeneity of mesothelioma or other malignancies with variable cellular composition. High yield and quality RNA was extracted from precision annotated, tumor-enriched subsamples prepared by combining individual microaliquots with the highest tumor cellularity estimates. Microaliquoting provides accurate cell content annotation and permits genomic analysis of enriched subpopulations of cells without fixation or amplification.
Subject(s)
Frozen Sections , Tissue Fixation , Adult , Aged , Aged, 80 and over , Female , Frozen Sections/methods , Humans , Male , Mesothelioma , Middle Aged , Pathology, Molecular , RNA/analysis , Tissue Fixation/methodsABSTRACT
Idiopathic thrombocytopenic purpura (ITP) is a relatively common autoimmune disorder among women of child-bearing age. It has a frequency of approximately one to two per 1,000 live births, accounting for about 3% of all cases of maternal thrombocytopenia at delivery. ITP in pregnancy necessitates the management of two patients, the mother and her baby; hence, the close collaboration of a multidisciplinary group composed of a hematologist, obstetrician, and pediatrician is essential. Our understanding of thrombocytopenia in pregnancy has evolved considerably over the last decade, yet the optimal diagnostic and treatment strategies for ITP in pregnancy continues to create controversy. In reviewing the recent literature, there is resurgence in the trend towards treating these patients in a more conservative fashion. This review will summarize the current approach to the diagnosis of ITP in pregnancy, as well as explore the pertinent and controversial issues of investigation and management.
Subject(s)
Pregnancy Complications, Hematologic/therapy , Purpura, Thrombocytopenic, Idiopathic/therapy , Diagnosis, Differential , Disease Management , Female , Humans , Infant, Newborn , Intracranial Hemorrhages/etiology , Male , Pregnancy , Pregnancy Complications, Hematologic/diagnosis , Pregnancy Complications, Hematologic/epidemiology , Purpura, Thrombocytopenic, Idiopathic/diagnosis , Purpura, Thrombocytopenic, Idiopathic/epidemiology , Thrombocytopenia/epidemiology , Thrombocytopenia/immunology , Thrombocytopenia/therapyABSTRACT
Equine airway smooth muscle is innervated by vagal efferents and, in addition, displays spontaneous mechanical activity. The preparation thus appears to contain at least two discrete excitable components, the cholinergic neural elements and the smooth muscle membrane. Indomethacin (INDO), a cyclooxygenase (CO) inhibitor, exerts a considerable potentiation of function in this preparation. The latter may be effected indirectly, through loss of the inhibitory effect of endogenous prostaglandin E2 (PGE2) on neural acetylcholine release and through direct effects on smooth muscle of the generally antagonistic CO and lipoxygenase (LO) metabolites. The present studies were designed to assess the relative contributions of altered arachidonic acid metabolism on those respective elements. The utility of the model, in terms of distinguishing neural and myogenic components, was assessed by examining the effects of the muscarinic antagonist atropine (ATR) and the neurotoxin tetrodotoxin (TTX) on the stimulus-response (SR) relationship. The observations that TTX and ATR produced similar rightward (but not downward) shifts of the SR curve and that D-600 inhibited the TTX-insensitive responses are consistent with a selective activation of the muscle by the nerves at lower voltages and a direct stimulation of the muscle at higher voltages. INDO potentiated both the neural and myogenic components of the SR curve, effects which were sensitive to ATR and 5,8,11,14-eicosatetraynoic acid, an inhibitor of LO, and reversed by PGE2. The finding that PGE2 at low doses (10(-8) M) inhibited responses at lower voltages and that at higher concentration (10(-7) M) it shifted the SR curve right and downward suggested that neurotransmitter release is more sensitive to PGE2 inhibition than is muscle response.(ABSTRACT TRUNCATED AT 250 WORDS)
