Digital image analysis of hematopoietic colonies in vitro

A system for automatic analysis of in vitro hematopoietic colonies is described and evaluated. With the standard resolution provided by video cameras, the improvement in visualization obtained using features other than size and darkness when classifying potential colonies appears to be limited. We confirmed this by comparing results obtained with the test system with those obtained with a commercial one. However, for some applications it may be useful to supplement the system with specific methods, e.g., to separate merged colonies. Digital image analyses provide new possibilities, for instance of measuring the total cellularity of the dish or analyzing colonies according to the size and cell density of each colony. Examples provided are time course studies of colony development, cellularity feedback effects on colony sizes, and bell-shaped dose-response curves for the growth stimulation obtained by certain conditioned media on a subpopulation of progenitor cells that gives rise to large colonies.     

Infants' detection of increments in low- and high-frequency noise

A visually reinforced operant paradigm was employed to examine the relationship between the difference limen (DL) for intensity and level of the standard during infancy. In Experiment 1, 7-month-old infants and adults detected increments in continuous noise presented via headphones at each of four levels ranging from 28 to 58 dB SPL. Noise stimuli were 2-octave bands centered at either 400 or 4000 Hz, and increments were 10 and 100 msec in duration. Infants' DLs were significantly larger than those of adult subjects and significantly larger for low- than for high-frequency stimuli. For the high-frequency noise band, infants' DLs were generally consistent with Weber's law, remaining essentially constant for standards higher than 28 dB SPL (3 dB SL) for 100-msec increments and 38 dB SPL (13 dB SL) for 10-msec increments. For low-frequency noise, infants' absolute thresholds were exceptionally high, and sensation levels of the standards were too low to adequately describe the relationship. In Experiment 2, 7-month-old infants detected 10- and 100-msec increments in 400-Hz noise stimuli presented in sound field. Infants' low-frequency DLs were large at low intensities and decreased with increases in level of the standard up to at least 30 dB SL. For both low- and high-frequency noise, the difference between DLs for 10- and 100-msec increments tended to be large at low levels of the standard and to decrease at higher levels. These results suggest that the relationship between the DL and level of the standard varies with both stimulus frequency and duration during infancy. However, stimulus-dependent immaturities in increment detection may be most evident at levels within approximately 30 dB of absolute threshold.     

5-Aminolevulinic acid induced endogenous porphyrin fluorescence in 9L and C6 brain tumours and in the normal rat brain

A new approach in photodynamic therapy is the use of endogenous porphyrins for sensitisation of tumours to light. The induction of endogenous porphyrins after intravenous injection of 5-aminolevulinic acid (ALA, 200 mg kg-1) was studied in 23 rats, bearing intracranial 9L or C6 tumours. After 0, 2, 4, 6, 8, and 22 hours the rats were sacrificed and the fluorescence distribution of endogenous porphyrins was studied in brain tissue sections with a standard fluorescence microscope and a confocal laser scanning microscope. The role of blood-brain barrier disruption on porphyrin production was studied in 2 rats with a cryo-lesion of the cortex. Additionally, 9L and C6 tumour cell cultures were incubated with ALA for 8 hours in vitro. Fluorescence was measured with a fluorescence spectrophotometer in cell cultures and in the brain sections. Porphyrins were detected in vitro in the tumour cells from 2 hours onwards and ex vivo in the tumour sections mainly from 2 to 8 hours, by 22 hours porphyrin fluorescence had almost disappeared. The contralateral brain showed low fluorescence levels between 2 and 6 hours after ALA administration. At the site of the cryo-lesions low fluorescence was measured 6 hours after ALA administration. The 9L tumours fluoresced homogeneously, with a sharp demarcation towards normal brain tissue. Fluorescence in the C6 tumours was patchy, with a poorly fluorescing edge. In both tumour models fluorescence was also detected in brain surrounding the tumour and sometimes in contralateral white matter and ventricle ependyma and pia mater. The slight increase of porphyrin fluorescence in the normal brain of tumour bearing rats, compared to the absence of this in rats without a tumour, was attributed to transport by bulk flow of porphyrins made in the tumours, and possibly also of circulating porphyrins or ALA leaking from the tumour vessels.     

Role of the TRAF binding site and NF-kappaB activation in Epstein-Barr virus latent membrane protein 1-induced cell gene expression

In this study, we investigated the induction of cellular gene expression by the Epstein-Barr Virus (EBV) latent membrane protein 1 (LMP1). Previously, LMP1 was shown to induce the expression of ICAM-1, LFA-3, CD40, and EBI3 in EBV-negative Burkitt lymphoma (BL) cells and of the epidermal growth factor receptor (EGF-R) in epithelial cells. We now show that LMP1 expression also increased Fas and tumor necrosis factor receptor-associated factor 1 (TRAF1) in BL cells. LMP1 mediates NF-kappaB activation via two independent domains located in its C-terminal cytoplasmic tail, a TRAF-interacting site that associates with TRAF1, -2, -3, and -5 through a PXQXT/S core motif and a TRADD-interacting site. In EBV-transformed B cells or transiently transfected BL cells, significant amounts of TRAF1, -2, -3, and -5 are associated with LMP1. In epithelial cells, very little TRAF1 is expressed, and only TRAF2, -3, and -5, are significantly complexed with LMP1. The importance of TRAF binding to the PXQXT/S motif in LMP1-mediated gene induction was studied by using an LMP1 mutant that contains alanine point mutations in this motif and fails to associate with TRAFs. This mutant, LMP1(P204A/Q206A), induced 60% of wild-type LMP1 NF-kappaB activation and had approximately 60% of wild-type LMP1 effect on Fas, ICAM-1, CD40, and LFA-3 induction. In contrast, LMP1(P204A/Q206A) was substantially more impaired in TRAF1, EBI3, and EGF-R induction. Thus, TRAF binding to the PXQXT/S motif has a nonessential role in up-regulating Fas, ICAM-1, CD40, and LFA-3 expression and a critical role in up-regulating TRAF1, EBI3, and EGF-R expression. Further, D1 LMP1, an LMP1 mutant that does not aggregate failed to induce TRAF1, EBI3, Fas, ICAM-1, CD40, and LFA-3 expression confirming the essential role for aggregation in LMP1 signaling. Overexpression of a dominant form of IkappaBalpha blocked LMP1-mediated TRAF1, EBI3, Fas, ICAM-1, CD40, and LFA-3 up-regulation, indicating that NF-kappaB is an important component of LMP1-mediated gene induction from both the TRAF- and TRADD-interacting sites.     

Noniatrogenic esophageal trauma

Few guidelines are available with which to facilitate treatment in patients with noniatrogenic injuries of the esophagus. Early diagnosis and proper management are essential if a good outcome is to be expected. In an effort to define better the treatment of patients with penetrating and blunt injuries of the esophagus, we report our recent 5-year experience at an urban trauma center. From July 1988 to June 1993, nineteen patients with esophageal perforations from penetrating (18) and blunt (1) trauma were identified by our trauma registry. There was no mortality in this group of patients and morbidity was mostly due to associated injuries. Eleven cervical esophageal injuries were repaired. One cervical injury was treated by stopping oral intake and giving intravenous antibiotics. The neck was not drained in 10 of the surgical cases. In 1 patient a tracheoesophageal fistula developed, which later was repaired with a pectoralis muscle flap. Seven perforations were identified in the thoracic (2) and abdominal (5) portions of the esophagus. All were due to gunshot wounds. In 4 cases, a fundal wrap was used to reinforce the repairs. Postoperative contrast studies confirmed that all repairs were intact. We conclude that penetrating and blunt tears of the esophagus can be repaired safely with minimal mortality. Morbidity is usually from associated injuries such as to the spinal cord and trachea. When identified early, cervical esophageal injuries do not need to be drained routinely.     

Some pharmacological studies on Artemisia herba-alba (Asso.) in rabbits and mice

This clinical research plan was designed to evaluate the predictable and current developments, growth, and stability of the economic status of part-time or full-time African-American practicing surgeons. In many communities throughout the United States, the economic successes of certified or experienced black surgical specialists are inhibited or modified by mistrust, jealousy, professional disengagements, prejudices, and self-hatred by black and white physicians and lay African Americans. Nevertheless, there are subtle and overt evidences of increasing satisfactory and above average associative interprofessional relationships between African-American physicians and surgical specialists. One hundred African-American surgical specialists from thirty-four communities in the United States were interviewed at periodic intervals over a period of three decades. Recommendations for improvements and changes are presented.     

A framework for the characterization and verification of embedded phase-locked loops

With the increasing use of phase-locked loops (PLLs) embedded in FPGAs, ASICs, and system-on-chip (SOC), there is a growing need for methods to verify their operation. This paper describes a general and organized list that includes tests for lock, jitter, stability, and modulation response. The list is offered as a guide to the verification and testing of an embedded PLL. For that reason, it is presented in such a way as to allow the reader to determine the extent to which the verification will be carried out. The tests are covered from the easiest to the most complicated to perform, with the amount of information gathered increasing along with the complexity of the test.     

Treatment of patients admitted to the hospital with congestive heart failure: specialty-related disparities in practice patterns and outcomes

OBJECTIVES: This study sought to define specialty-related differences in the care and outcome of patients admitted to the hospital with congestive heart failure (CHF). BACKGROUND: Congestive heart failure is the leading diagnosis-related group (DRG) discharge diagnosis in the United States and accounts for an estimated annual hospital cost in excess of $7 billion. The clinical impact of aggressive CHF management and the importance of the subspecialist in guiding this care have not been evaluated. METHODS: To define differences in physician practice patterns, we performed a chart review of consecutive patients admitted to a university teaching hospital with a primary DRG discharge diagnosis of CHF. We compared treatment and outcome of patients cared for by a generalist (n = 160) and those whose care was guided by a cardiologist (n = 138) during their index hospital period with CHF and over the next 6 months. RESULTS: At our institution, > 50% of patients admitted to the hospital with CHF cared for by generalists alone had minimal (New York Heart Association functional class I or II) symptoms, compared with < 15% of those cared for by a cardiologist (p < 0.01). Although generalists' patients underwent significantly fewer in-hospital diagnostic tests and had shorter lengths of stay, they had a 1.7-fold increased risk of readmission for CHF within 6 months (p < 0.05). Six-month cardiac and all-cause mortality were not significantly different between the groups. The type of physician caring for the patient and a history of diabetes, previous CHF or myocardial infarction were independent predictors of readmission for CHF. CONCLUSIONS: Involvement of a cardiologist in the care of patients admitted to the hospital with CHF is associated with increased use of diagnostic testing, longer hospital stays and improved clinical outcome. These results substantiate practice guidelines that suggest a role for cardiologists in the care of symptomatic patients with CHF.