Filamentous phage displaying the extracellular domain of the hLH/CG receptor bind hCG specifically.

We have expressed the extracellular binding domain of the human LH/CG receptor as a fusion with the cpIII filamentous phage coat protein. These fusion phage are able to specifically bind ELISA plates coated with hCG. Preincubation of the phage with hCG before exposure to the coated plates inhibited binding of phage, whereas human FSH had no effect. Furthermore, addition of anti-hLH/CG receptor antisera inhibited binding of phage to the plates. We have also tested the effect of monoclonal antibodies to hCG on phage binding. Monoclonal antibodies that can bind hCG when it is bound to native receptor do not inhibit phage binding. These include B105 and A109 Alternatively, B107 and B109 have an inhibitory effect on phage binding. They cannot bind hCG when it is bound to receptor and are likely directed against epitopes at or near the receptor binding interface. Therefore, these fusion phage exhibit the same binding specificity as the wild-type receptor and likely display the extracellular domain of the hLH/CG receptor on their surface in the native conformation. Phage display is a potentially useful tool for studying the hLH/CG receptor structure and in screening for synthetic compounds that compete with hormone for receptor binding.

Increased serum concentrations of cholesterol and triglycerides in the progression of breast cancer.

The course of serum concentrations of cholesterol and triglycerides was investigated in patients with advanced breast cancer. The patients studied were divided into two groups according to their clinical status: group-I consisted of 51 patients who already had metastases at the start of the investigation, but progressed further during the time of observation; group-II consisted of 14 patients in remission who experienced recurrence of disease while under observation. In group-I, 28 patients (54.9%) were found to have normal serum triglyceride levels at the beginning of the observation period; 22 patients (78.6%) from this group experienced a significant (P less than 0.0001) increase above the normal range upon further disease progression. Similarly, serum cholesterol levels were normal in 32 patients (62.8%) at the start of the investigation, but increased significantly (P less than 0.0001) above the normal range upon disease progression. In group-II, 8 patients (57.2%) had normal serum triglyceride levels at the beginning of the observation period, but the levels increased in 4 patients (50%) significantly (P less than 0.005) upon the occurrence of metastases. Within the same group, a significant increase (P less than 0.001) of initially normal serum cholesterol levels was found in 4 (44.9%) out of 9 patients. In summary, a rise in serum levels of triglycerides and/or cholesterol should receive increased attention and could indicate progression or recurrence of breast cancer.

Value measurement in health care: a new perspective.

Vital to the success of any healthcare organization is the ability to obtain useful information and feedback about its performance. In particular, healthcare organizations need to begin to understand how non-value-adding work activities detract from their bottom lines. Additionally, financial managers and information systems need to provide data and reports throughout the continuum of care. Overall, healthcare organizations must align the management information and control systems with the planning and decision-making processes. The horizontal information system is a tool to manage three common problems facing today's healthcare managers: (1) the use of existing information to focus on control rather than improve business, (2) failure to focus on satisfying the customer, and (3) failure to combine their efforts with those of the employees by developing trust and a common focus.

A randomized phase II trial using two different treatment schedules of gemcitabine and carboplatin in patients with advanced non-small-cell lung cancer.

BACKGROUND: Gemcitabine and carboplatin combination therapy is an active and tolerable regimen in the treatment of non-small-cell lung cancer (NSCLC). Twenty-eight- and 21-day regimens without day-15 administration of gemcitabine are common; however, it remains unclear which offers the optimal therapeutic index. METHODS: This trial evaluated two schedules of the combination of gemcitabine and carboplatin: gemcitabine (1100 mg/m on days 1 and 8) plus carboplatin (area under the curve = 5 on day 8) every 28 days, or gemcitabine (1000 mg/m on days 1 and 8) plus carboplatin (area under the curve = 5 on day 1) every 21 days. Eligible patients in this trial had stage IIIB (with malignant pleural effusion) or stage IV NSCLC with no prior chemotherapy. The primary objective was to evaluate progression-free survival, with secondary objectives of overall survival, response rate, and toxicity. RESULTS: One hundred patients were randomized and enrolled from October of 2000 to January of 2002 into this multi-institutional study (48 for the 28-day regimen and 52 for the 21-day regimen). Baseline demographics were well matched, and a majority of patients (85%) enrolled with stage IV disease. Median progression-free survival and response rates were 3.8 months and 22.9%, respectively, with the 28-day regimen, and 4.9 months and 40.4%, respectively, with the 21-day regimen. Median survival was 8.7 months with the 28-day regimen and 8.0 months for the 21-day regimen. One- and 2-year survival rates were 34.7% and 8.7%, respectively, with the 28-day regimen, and 36.5% and 16.8%, respectively, with the 21-day regimen. Differences in progression-free survival (log-rank statistic, p = 0.5786), response rate (Fisher's exact test, p = 0.0859) and overall survival (log-rank statistic, p = 0.3568) were not statistically significant. Grade 3 to 4 hematologic toxicities occurred with a greater frequency in the 21-day regimen. No grade 3 to 4 nonhematologic toxicity (except nausea/vomiting with the 28-day regimen) was observed in more than 10% of patients in either treatment arm. CONCLUSION: Gemcitabine plus carboplatin is active and well tolerated in advanced NSCLC. Both regimens may be considered for further study. Although the 21-day regimen appeared to be associated with preferable outcomes, differences between treatment groups were not statistically significant.

Effects of tactile and kinesthetic stimulation on premature infants.

Forty premature infants, divided equally into matched treatment and control groups, were studied to determine the effect of compensatory tactile and kinesthetic stimulation on clinical course. The treatment group received a 15-minute period of tactile and kinesthetic stimulation once daily on Days 1-10 postpartum; control group received routine care. Treated infants showed increased stooling frequency on Days 5-10, and increased feeding intake on Days 6-10. Treated infants gained increasingly more weight each day, but these differences did not reach statistical significance. These results suggest that single daily stimulation treatments in the first 10 days postpartum were sufficient to enhance the clinical course of premature infants.

Pulmonary manifestations of Waldenstrom macroglobulinemia.

Waldenstrom macroglobulinemia (WMG), a proliferation of malignant monoclonal IgM secreting plasmacytoid lymphocytes in lymph nodes, spleen, and marrow, usually pursues a chronic clinical course. A patient with WMG for five years who developed pulmonary tumors consisting of plasmacytoid lymphocytes prompted a review of the literature for pulmonary manifestations of WMG. Twenty-six males and 18 females, ranging in age from 33 to 84 years, have been reported with histologically proven pulmonary involvement by WMG. The x-ray findings, evident in most patients when first seen, consisted of masses (22 patients), inflitrates (31 patients), and pleural effusions (19 patients). Most patients (24) had two or more of these manifestations but only five, in addition to our patient, had isolated pulmonary nodules. Isolated pulmonary infiltrates were found in ten patients and isolated pleural effusions in only four. Symptoms at the onset of pulmonary involvement included dyspnea (54%), nonproductive cough (33%), and chest pain (7%); 15% were asymptomatic. Pulmonary manifestations, like other features of WMG, respond to alkylating agents or irradiation and do not appear to affect prognosis adversely. Pulmonary involvement should be suspected in any patient with WMG who develops an abnormal chest x-ray.

Granule enzymes of polymorphonuclear neutrophils: A phylogenetic comparison.

The functional significance of granule enzymes in polymorphonuclear leukocytes (PMN) is not fully understood because of the multiplicity of the enzymes and the rare occurrence of deficiencies in man. In order to select appropriate laboratory animals for functional studies, a phylogenetic comparison of enzyme levels in animal and human PMN was undertaken. Neutrophils were obtained from a variety of laboratory animals and man; the activities of alkaline phosphatase, lysozyme, myeloperoxidase, and beta-glucuronidase were determined by histochemical and analytical techniques. Marked interspecies differences in enzyme activity were found; many species were deficient in alkaline phosphatase or lysozyme. Differences in pH optima and metal requirements of alkaline phosphatase were not of sufficient magnitude to explain the variations of this enzyme.

Characterization of monkey peripheral neutrophil granules during infection.

Rhesus monkey (Macaca mulatta) neutrophils were shown to contain the azurophilic granule maker enzymes myeloperoxidase and beta-glucuronidase but were deficient in the specific granule markers alkaline phosphatase (AKP) and lysozyme. Isopycnic centrifugation of leukocyte homogenates on linear sucrose gradients resulted in cosedimentation of myeloperoxidase and beta-glucuronidase with an equilibrium density of 1.18. After an intravenous inoculation of monkeys with Salmonella typhimurium AKP activity became marked, whereas that of beta-glucuronidase decreased and myeloperoxidase remained unchanged. Lysozyme was undetected throughout the course of the experiment, but was present in oil-induced peritoneal macrophages and peripheral mononuclear cells. The induced AKP exhibited partial latency and had an equilibrium density of 1.15. It is unclear, however, whether the induced AKP is associated with specific granules or cytoplasmic membranes. Hence, while these data are consistent with the presence of azurophilic granules in polymorphonuclear neutrophils from infected monkeys, the presence of specific granules in polymorphonuclear neutrophils of both uninfected and infected monkeys remains moot.

[Differential diagnosis and therapy of bradycardic arrhythmias]. / Differentialdiagnose und Therapie bradykarder Rhythmusstörungen.

The most important symptoms in bradycardia are vertigo, dizziness and syncopy due to diminished cerebral blood sypply. Cardial symptoms are cardiac insufficiency and angina pectoris. By means of ECG, especially Holter-ECG, carotid sinus massage, atropin test and invasive methods (atrial stimulation, His-bundle ECG) sinu-nodal dysfunction, carotid sinus syndrome, bradyarrhythmia absoluta and AV-block can be diagnosed. Pharmacological treatment is only useful in acute situations. For symptomatic bradyarrhythmias the implantation of a Pacemaker is the therapy of choice. Individual treatment of the various types of bradyarrhythmia and the patients special needs is possible through the evolution of pacemaker technology.

Immunocytochemical identification of azurophilic and specific granule markers in the giant granules of Chediak-Higashi neutrophils.

We used immunofluorescent microscopy to characterize the abnormal granules in neutrophils from five patients with Chediak-Higashi disease. Monospecific antiserums to the azurophilic markers myeloperoxidase, elastase, cathepsin G and lysozyme, and to the specific granule markers lactoferrin and lysozyme, were labeled with fluorescein and rhodamine and were used to demonstrate two antigens in the same cell simultaneously. The abnormal granules in Chediak-Higashi neutrophils contained both azurophilic and specific granule markers. Normal-appearing lactoferrin-positive granules were also present, but normal azurophilic granules were not seen. Analysis of bone-marrow samples from two of these patients suggested that the abnormal granules were formed during granulocyte maturation by the progressive aggregation and fusion of normally formed azurophilic and specific granules. These results are consistent with a membrane abnormality or a defect of microtubular function leading to inappropriate granule fusion, and suggest that the granular abnormality is more generalized than previously appreciated.

Ultrasonographically guided thyroid biopsy: a review with emphasis on technique.

We describe our technique for ultrasonographically guided fine-needle aspiration biopsy of the thyroid that achieves a high rate of diagnostic specimens. Indications for ultrasonographically guided fine-needle aspiration biopsy included a difficult-to-palpate thyroid nodule and previously unsuccessful palpation-guided fine-needle aspiration. Ultrasonographically guided fine-needle aspiration biopsy was performed on 316 thyroid nodules in 306 patients. Adequate cytologic specimens were obtained in 97.2% of the nodules in which biopsy was performed, with a 2.8% rate of inadequate cellularity. Two helpful aspects of this technique that were thought to improve the overall diagnostic yield were the use of color and power Doppler "vascular mapping" of the nodule just before biopsy and on-site cytologic control.

Pulsed versus continuous wave excitation mechanisms in photodynamic therapy of differently graded squamous cell carcinomas in tumor-implanted nude mice.

The effectiveness of photodynamic therapy (PDT) in vivo was compared between the pulsed excimer laser-pumped dye laser system (EDL) and the continuous wave (cw) argon laser-pumped dye laser system (ADL). Serial subcutaneous transplantation was used to implant thymus aplastic nude mice with different grades of malignancy of two human squamous cell carcinomas (SCCs). Forty-eight hours after i.v. injections of a hematoporphyrin derivative (Photosan 3), the animals were irradiated with either pulsed-EDL or cw-ADL laser light at a tumor depth of 4-5 mm. The irradiation data were chosen as follows: EDL and ADL wavelength 630 nm, total dose 150 J/cm2, irradiation time 27.78 min; EDL repetition rate 30 Hz, single pulse energy 3 mJ, pulse width 20 ns; ADL intensity 90 mW/cm2. The effects of PDT were studied either by long-term observation of the animals treated or by evaluation of hematoxylin-eosin and Ki-67 histological sections of tumors 48 h after treatment. The EDL system proved to be at least as efficient as the ADL system as judged by the number of complete remissions. This became particularly evident in the treatment of the lower-graded tumors, with a good response observed in both transplanted SCCs. However, the higher-graded tumors showed a better response to PDT.

Carbamazepine levels in breast milk.

The carbamazepine (CBZ) concentration of breast milk was determined in 19 epileptic patients of whom 13 were receiving CBZ monotherapy. Two lactation periods under CBZ monotherapy were followed in 2 of these 13 patients. The CBZ concentration of the breast milk was 36.4% of the maternal CBZ serum level (mean value). The correlation coefficient was 0.44; the regression line follows the equation y = 0.12X + 1.63. The CBZ concentration was measured in a total of 50 milk samples during the lactation period (cases of monotherapy plus combination therapy). The CBZ concentration ranged from 1 to 4.8 micrograms/ml (mean 2.5 micrograms/ml). The free CBZ concentration in the maternal serum was measured in four patients. The correlation between CBZ in the breast milk and free CBZ levels was 1.6 to 1.8. The ratio of carbamazepine-epoxide (ECBZ) in the milk to ECBZ determined in serum was 0.53. Continuing galactorrhea after delivery was observed in one patient. The CBZ and ECBZ levels determined 3.5 years after delivery were higher in the milk than in the serum. The observations of breast-fed and non-breast-fed children whose mothers were on CBZ monotherapy disclosed poor suckling in only 1 of 15 infants.

Malignant histiocytosis: a cytochemical and electron microscopic study of an unusual case.

A 25-year-old black female presented with lymphadenopathy, fever and anemia of two months duration. The diagnosis of malignant histiocytosis was made on the basis of histiocytic infiltrations in the sinuses of spleen, liver and lymph nodes and by the demonstration of erythrophagocytosis in bone marrow. Following splenectomy, the patient developed a leukemic phase with as many as 50 X 10(9) abnormal histiocytes/l and bone marrow necrosis. This patient was also atypical because of multiple granulomas in liver, spleen and lymph nodes. Cytochemical and immunofluorescent stains confirmed that the abnormal cells were derived from the monocyte-macrophage series. Electron microscopy was used to further characterize this abnormal cell population. The electron microscopic and cytochemical evidence confirms that the malignant cells in malignant histiocytosis are derived from monocytes.

Immunocytochemical identification of abnormal polymorphonuclear neutrophils in patients with leukemia.

High titer, monospecific antibodies to human granulocyte myeloperoxidase, cathepsin G, elastase, lysozyme, and lactoferrin were conjugated with fluorescein and rhodamine and used for immunofluorescent staining of mature neutrophils obtained from 25 patients with acute and chronic leukemia. In 11 (44%) of the patients, two populations of mature neutrophils were detected. The abnormal cells were identified by complete deficiency of one or more markers and constituted 10%-100% of the total number of neutrophils. This immunocytochemical approach may permit recognition of mature cells derived from leukemic clones, and serial determinations of the ratio of normal to abnormal cells may be useful in the management of patients with leukemia.

Further evidence for linkage of low-mid frequency hearing impairment to the candidate region on chromosome 4p16.3.

We have investigated a three-generation family with an autosomal dominant low-mid frequency hearing loss. Audiograms show consistently a hearing threshold of 50+/-20 db hearing loss (HL) between 250 Hz and 1-2 kHz. Normal hearing level was reached between 3 and 6 kHz in all examined children. Adult patients show an additional hearing impairment (HI) in the mid and higher frequencies that seems to differ from presbyacusis. The HI is always bilateral and symmetrical. Genes causing non-syndromic autosomal-dominant deafness with HI in the low and mid frequencies were previously mapped to chromosome 4p16.3 (DFNA6, DFNA14) and chromosome 5q31 (DFNA1). After exclusion of linkage to DFNA1 on chromosome 5, we mapped the candidate gene region to the DFNA14 and DFNA6 loci, between the genetic markers D4S432 and D4S431, located on chromosome 4. This is a further family in which evident linkage of low-mid frequency HI to the candidate region on chromosome 4p16.3 has been found.