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At least 433 records · Page 24Linked to original sources
PubMed1993

Antitumor response to recombinant murine interferon gamma correlates with enhanced immune function of organ-associated, but not recirculating cytolytic T lymphocytes and macrophages.

The mechanism of therapeutic activity for recombinant murine interferon-gamma (rMu IFN gamma) in the treatment of metastatic disease was investigated by comparing effector cell augmentation with therapeutic activity in mice bearing experimental lung metastases (B16-BL6 melanoma). Effector cell functions in spleen, peripheral blood, and lung (the tumor-bearing organ) were tested after 1 week and 3 weeks of rMu IFN gamma administration (i.v. three times per week). Natural killer (NK), lymphokine-activated killer (LAK), cytolytic T lymphocyte (CTL) activities against specific and nonspecific targets, and macrophage tumoristatic activity were measured. rMu IFN gamma demonstrated immunomodulatory activity in most assays of immune function. The optimal therapeutic protocol of rMu IFN gamma (2.5 x 10(6) U/kg, three times per week) prolonged survival and decreased the number of pulmonary metastatic foci. This therapeutic activity was correlated with specific CTL activity from pulmonary parenchymal mononuclear cells (PPMC), but not from spleen or blood. Macrophage tumoristatic activity in PPMC also correlated with therapeutic activity, but activity in alveolar macrophages did not. However, therapeutic activity did not correlate with NK or LAK activity at any site. These results demonstrate that the optimal therapeutic protocol is the same as the optimal immunomodulatory dose for pulmonary CTL and macrophage activities. Furthermore, while immunological monitoring may help to optimize treatment protocols, current monitoring procedures that use readily accessible sites, particularly peripheral blood, may not accurately predict the therapeutic efficacy of biological response modifiers in clinical trials.

Animals↗
PubMed2001

Development of an optimized interaction-mating protocol for large-scale yeast two-hybrid analyses.

BACKGROUND: Protein-protein interactions have decisive roles in almost all aspects of the structural and functional organization of cells. But in spite of the increasing amount of complete genome sequence data, the ability to predict protein function from sequences alone is limited. Therefore comprehensive analysis of protein-protein interactions, as derived from the yeast two-hybrid mating system, will yield valuable information for functional biology on a proteomic scale. RESULTS: We have developed an optimized interaction mating protocol for the yeast two-hybrid system, which gives increased mating efficiencies. This significantly reduces the effort and cost of cDNA library screening and allows multiple parallel approaches. Improved preincubation conditions before mating, and optimal cell densities and cell ratios enable almost quantitative mating of the yeast cells carrying the cDNA library. We have proved the applicability of this technology using 20 bait proteins to screen an Arabidopsis thaliana cDNA library, in spite of bait-dependent variations in mating efficiency. CONCLUSIONS: The improved yeast two-hybrid interaction-mating protocol presented here allows the multiple parallel screening of cDNA libraries. It can be carried out without specialized equipment and has the potential to be standardized and automated.

Arabidopsis↗
PubMed2000

[An optimal follow-up protocol for renal cell carcinoma based on the occurrence of recurrences after surgery].

INTRODUCTION AND OBJECTIVE: The tumor biology of renal cell carcinoma (RCC) remains poorly understood and the prognosis of which is unpredictable. To define an optimal follow-up protocol in nonmetastatic RCC, tumor recurrences and the clinical course were assessed in the patients who had undergone nephrectomy. PATIENTS AND METHODS: Between March 1972 to July 1999, 265 patients with PT1-3N0 (Nx) M 0 RCC underwent radical or partial nephrectomy. Clinicopathological variables were compares with the time of the first recurrence, site of metastasis and reason for diagnosis. RESULTS: In July 1999, the median (range) follow-up for the surviving patients was 50 (1-244) months. Metastases were diagnosed in 45 of the 265 patients (17%). Sixty percent of the patients had their metastases diagnosed within 3 years and 87% by 6 years. The incidence and the mean time to first diagnosis of recurrence were 9.0% and 43.6 months for pT1, 32.4% and 39.6 months for pT2, and 32.7% and 25.0 months for pT3. Metastatic sites and the mean time of diagnosis were 40.6 months in lung (46.7%), 26.1 months in bone (17.8%), 18.6 months in retroperitoneal organs (11.1%), 38.0 months in liver (6.7%), 51.0 months in thyroid gland (4.4%) and 31.0 months in brain (2.2%). Most recurrence in lung, retroperitoneal organs and liver were incidentally diagnosed, whereas recurrences in bone, thyroid gland and brain had some symptoms associated with metastases. Incidence of lung metastasis in pT1 (9 patients, 56%) and pT2 (6, 55%) was greater than that in PT3 (6, 33%). In contrast, incidence of abdominal metastasis (retroperitoneal organs and the liver) in pT3 (4 patients, 33%) was grater than pT1 (2, 12%) and PT2 (1, 16%). CONCLUSIONS: The results indicate that the risk of tumor progression and site of metastasis seemed to depend on original TNM stage. Therefore stage specific postoperative follow-up protocol should be considered. The yearly follow-up may suffice for pT1a patients after surgery. For pT1b and pT2 patients, the follow-up should be intense during the first 3 years by a routine chest X-ray. For pT3 patients, the follow-up during the first 2 years may be important. Particularly, considering the high incidence of abdominal metastasis in these patients, examination by CT scan is recommended every 6 months in the first 2 years. No need for bone scintigraphy and brain CT was indicated for asymptomatic patients.

Carcinoma, Renal Cell↗
PubMed1986

Dissociation of therapeutic and toxic effects of polyinosinic-polycytidylic acid admixed with poly-L-lysine and solubilized with carboxymethyl cellulose in tumor-bearing mice.

In this paper, we describe a study of the therapeutic parameters (dose and schedule) and immunomodulatory activity (macrophage, natural killer cell, and T-cell number and function) of polyinosinic-polycytidylic acid admixed with poly-L-lysine and solubilized with carboxymethyl cellulose [poly(I,C)-LC] in the treatment of MBL-2 tumor ascites. Tumor-bearing mice received an optimal therapeutic protocol [100 micrograms poly(I,C)-LC administered twice a wk], a maximum tolerated dose [50 micrograms poly(I,C)-LC administered daily], or the optimal immunomodulatory protocol for normal mice [10 micrograms poly(I,C)-LC administered daily]. The percentage of tumor-associated macrophages and their cytotoxic activity correlated with host survival. In addition, splenic T-cell activity correlated with host survival, and splenic natural killer cell function had a near significant correlation with host survival. These results indicate that the optimal dose and schedule of poly(I,C)-LC for immunomodulation in tumor-bearing animals are also the optimal therapeutic protocol but have less toxicity than the maximum tolerated dose.

Animals↗
PubMed2003

Protocol to determine the optimal intraoral passive shim for minimisation of susceptibility artifact in human inferior frontal cortex.

At higher static magnetic field (B(0)) strengths (>/=3 T), the study of human inferior frontal cortex (IFC) when utilising a variety of MRI techniques is severely hampered by the presence of susceptibility artifacts. This is particularly the case for blood oxygenation level-dependent functional MRI, where large signal voids are generally encountered in the frontal lobes. A previous study described an approach to artifact correction involving a mouth insert consisting of a prototype diamagnetic passive shim [Magn. Reson. Med. 48 (2002), 906]. Here we extend that method by investigating the effect of five different intraoral passive shims on B(0) homogeneity and echoplanar imaging susceptibility artifacts within the brain, and particularly the IFC, of six subjects. The optimal passive shim is shown to be subject- and study-specific, providing an average reduction in mean absolute B(0) offset within the IFC of 57%, along with a concomitant reduction in echoplanar susceptibility artifact. All subjects were at ease while wearing the intraoral shims. A 4-min in vivo protocol to determine the optimal passive shim from the available set, utilising intrinsic structural and B(0) subject data, is described and shown to be accurate and reliable.

Artifacts↗
PubMed1991

Weaning of three hybridoma cell lines to serum free low protein medium.

A general weaning procedure is described which allowed a range of hybridomas to be weaned readily off serum without loss of antibody production. Initial work was carried out with one cell line only (SPO1 cells) and one serum substitute containing a final protein concentration of 40 mg l-1. The SPO1 cells were first adapted to a range of readily available basal media and then weaned off serum by a range of protocols. From this work an optimal weaning protocol and basal medium for weaning were determined. These were then used to wean the SPO1 cells and two other cell lines off serum with a second, protein free, serum substitute with varying concentrations of defined proteins added. All three cell lines investigated were readily weaned off serum by this protocol at protein concentrations as low as 1 mg l-1. No loss of antibody production was observed with any of the cell lines. The weaning procedure outlined in both simple and rapid and has been successfully adopted in our laboratory by relatively inexperienced cell culture technicians.

Animals↗
PubMed2003

Electroporation: theory and methods, perspectives for drug delivery, gene therapy and research.

Electroporation designates the use of short high-voltage pulses to overcome the barrier of the cell membrane. By applying an external electric field, which just surpasses the capacitance of the cell membrane, transient and reversible breakdown of the membrane can be induced. This transient, permeabilized state can be used to load cells with a variety of different molecules, either through simple diffusion in the case of small molecules, or through electrophoretically driven processes allowing passage through the destabilized membrane--as is the case for DNA transfer. Initially developed for gene transfer, electroporation is now in use for delivery of a large variety of molecules: From ions to drugs, dyes, tracers, antibodies, and oligonucleotides to RNA and DNA. Electroporation has proven useful both in vitro, in vivo and in patients, where drug delivery to malignant tumours has been performed. Whereas initial electroporation procedures caused considerable cell damage, developments over the past decades have led to sophistication of equipment and optimization of protocols. The electroporation procedures used in many laboratories could be optimized with limited effort. This review (i) outlines the theory of electroporation, (ii) discusses factors of importance for optimization of electroporation protocols for mammalian cells, (iii) addresses particular concerns when using electroporation in vivo, e.g. effects on blood flow and considerations regarding choice of electrodes, (iv) describes DNA electrotransfer with emphasis on use in the in vivo setting, and (v) sums up data on safety and efficacy of electroporation used to enhance delivery of chemotherapy to tumours in cancer patients.

Animals↗
PubMed1999

Protocols and future directions in imaging of renal artery stenosis: CT angiography.

Computed tomographic angiography (CTA) has become an established technique for minimally invasive imaging of renal artery stenosis. This article describes in detail the current scan protocols suggested for renal artery stenosis and explains the rationale behind the choice of such parameters as collimation, pitch, reconstruction interval, scan duration and scan length. It discusses the optimized use of contrast agents including bolus triggering and a saline flush to improve contrast agent utilization. Examples of optimized scan protocols are given for various scanner types including subsecond scanners and multislice computed tomography. With standard 1-s scanners, a slice collimation of 2 mm is recommended. A pitch factor of 1.5-2 is important for optimum volume coverage. Smoothing reconstruction kernels improve signal-to-noise ratio and allow for dose reduction. For optimum results, the plateau phase of contrast enhancement has to be matched as precisely as possible. Thus, a test bolus injection or an automated bolus triggering technique should be employed. Axial CT images remain the basis for making the diagnosis but require interactive viewing on a monitor display. Image processing (multiplanar reformats; maximum intensity projections; shaded surface displays) is employed to display the renal arteries. Given optimized examination protocols, high sensitivity and specificity can be achieved (90-98%) with negative predictive values of greater than 95%. Thus, a normal CT angiogram virtually rules out renal artery stenosis.

Angiography↗
PubMed1996

Multiparameter flow cytometric characterization of epidermal cell suspensions prepared from normal and hyperproliferative human skin using an optimized thermolysin-trypsin protocol.

Reliable flow cytometric analysis of normal and diseased skin requires pure epidermal single-cell suspensions. Several methods to separate the dermis from the epidermis are available. The proteolytic enzyme thermolysin separates the epidermis from the dermis at the lamina lucida and therefore permits reliable dermoepidermal separation. In the present study an optimized cell isolation procedure using thermolysin and trypsin is described, which is particularly suitable for punch biopsies. A 16-20-h (overnight) incubation of biopsies taken from normal and hyperproliferative skin with thermolysin (0.5 mg/ml) at 4 degrees C produced a selective separation of the dermis and epidermis. After a 30-min trypsin incubation (0.25 mg/ml) at 37 degrees C a cell suspension was produced which was characterized by minimal cell damage (cellular debris and clumps), a high recovery of basal cells and high quality DNA histograms. Furthermore, dermal contamination was very low. The thermolysin-trypsin separation methodology followed by triple-labelling flow cytometry provided a precise quantification of the percentage of keratin 10-positive cells, vimentin-positive cells and cells in S and G2M phases. Proliferative activity was selectively measured in the basal, the suprabasal and the non-keratinocyte compartment at various time intervals during epidermal regeneration after adhesive tape stripping. In contrast to the non-keratinocytes, the percentage of cells in S and G2M phases in the basal keratinocytes and in the suprabasal compartment increased 44-48 h after stripping. The increased proliferation following tape stripping was paralleled by an increased invasion of vimentin-positive cells into the epidermis and preceded by a decreased number of keratin 10-positive cells. Thermolysin-trypsin separation followed by three-colour flow cytometry permits a highly selective characterization of normal and hyperproliferative epidermis.

Adolescent↗
PubMed1999

Radiation dose for pedicle screw insertion. Fluoroscopic method versus computer-assisted surgery.

STUDY DESIGN: Comparison of the radiation dose between the traditional fluoroscopic approach and computed tomography (CT)-based computer-assisted surgery for pedicle screw placement was determined. OBJECTIVES: To evaluate the radiation dose delivered by fluoroscopy-controlled pedicle screw placement versus insertion guided by computer. To define the CT computer-assisted protocol, involving lower radiation exposure for the patient, that still provides acceptable image quality. SUMMARY OF BACKGROUND DATA: There are no published data describing the dose delivered in CT-based image-guided surgery, and there are few studies in which the organ dose and the effective dose delivered during pedicle screw insertion that is performed traditionally with fluoroscopic control are described. METHODS: Dose measurements were performed on two types (REMAB and RANDO) of anthropomorphic phantoms. Thermoluminescent dosimeters were used to measure the organ dose. Both phantoms were exposed to the fluoroscopic x-ray beam. The representative intraoperative scenario was determined by observation of 20 consecutive surgical interventions featuring pedicle screw implantation. For the CT dose measurement only, the REMAB phantom was used with two types of CT scanners. Three scanning protocols were evaluated: sequential, spiral optimized, and sequential optimized. Optimization of the scanning protocol included changes of anode current. The CT images were subsequently processed to achieve three-dimensional reconstruction of the lumbar spine for the computer-assisted intervention. RESULTS: Organ and effective doses were higher in any of the CT examinations than in the fluoroscopic procedure. There was a slight difference between doses registered during optimized spiral scanning and doses in the calculated optimized sequential CT protocol. Optimized sequential scanning was associated with an effective dose 40% lower than that in nonoptimized sequential scanning. The small anatomic structures of the spine could be easily recognized on each of the three-dimensional reconstructions, and all of them were suitable for use in computer-assisted surgery. CONCLUSIONS: Percutaneous pedicle screw insertion in the lumbar region of the spine, performed using fluoroscopic control, requires a lower radiation dose than do CT scans necessary for computer-assisted surgery. The CT radiation dose can be significantly decreased by optimization of the scanner settings for computer-assisted surgery. The advantages of computer-assisted surgery justify CT scans, when based on correctly chosen indications.

Adult↗
PubMed1996

Optimization of immunofluorescence methods by quantitative image analysis.

There is a growing trend towards the objective quantification of immunohistochemical staining. However, quantification has not been used previously to optimize the original published immunohistochemical methods. We present a quantitative method for analyzing immunofluorescence staining employing the Applied Imaging MAGISCAN image analysis system, which has then been used to optimize major aspects of the standard immunofluorescent staining protocols. The optimization process resulted in a method that increased specific staining up to fivefold over typical published protocols, with no increase in nonspecific staining. The method is extremely reproducible. For slides stained by a single experimenter in one batch on one day, the coefficient of variation between replicate means is 1.2%. The image analysis protocol gave a linear response with increasing antigen concentration, as determined by using purified antigen dried onto slides. The revisions to the standard protocol presented here can also be applied to nonquantitative staining. It will help users of immunofluorescence to maximize their staining and may enable the detection of previously undetected antigens.

Actins↗
PubMed2025

Synthetic community Hi-C benchmarking provides a baseline for virus-host inferences.

Microbiomes influence diverse ecosystems, and viruses increasingly appear to impose key constraints. While viromics has expanded genomic catalogs, host identification for these viruses remains challenging due to the limitations in scaling cultivation-based approaches and the uncertain reliability and relative low resolution of in silico predictions - particularly for understudied viral taxa. Towards this, Hi-C proximity ligation uses sequenced, cross-linked virus and host genomic fragments to infer virus-host linkages and has now been applied in at least ten studies. However, its accuracy remains unknown. Here we assess Hi-C performance in recovering virus-host interactions using synthetic communities (SynComs) composed of four marine bacterial strains and nine phages with known interactions and then apply optimized bioinformatic protocols to natural soil samples. In SynComs, standard Hi-C sample preparations and analyses showed poor normalized contact score performance (26% specificity, 100% sensitivity, incorrect matches up to class level) that could be dramatically improved by Z-score filtering (Z ≥ 0.5, 99% specificity), though at reduced sensitivity (62% down from 100%). Detection limits were established as reproducibility was poor below minimal phage abundances of 105 PFU/mL. Applying optimized bioinformatic protocols to natural soil samples, we compared virus-host linkages inferred from proximity-ligated Hi-C sequencing with predictions generated by in silico homology-based and machine learning-based bioinformatic approaches. Prior to Z-score thresholding, agreement was relatively high at the phylum to family levels (72%), but not at the genus (43%) or species (15%) levels. Z-score thresholding reduced sensitivity (only 34% of predictions were retained), with only modest improvements in congruence with bioinformatic methods (48% or 18% at genus or species levels, respectively). Regardless, this led to 79 genus-level-congruent virus-host linkages and 293 new ones revealed by Hi-C alone - i.e., providing many new virus-host interactions to explore in already well-studied climate-critical soils. Overall, these findings provide empirical benchmarks and methodological guidelines to improve the accuracy and reliability of Hi-C for virus-host linkage studies in complex microbial communities.

Genomics↗
PubMed2025

Benchmarking with synthetic communities provides a baseline for virus-host inferences from Hi-C proximity linking.

Microbiomes influence diverse ecosystems, and viruses increasingly appear to impose key constraints. While viromics has expanded genomic catalogs, host identification for these viruses remains challenging due to the limitations in scaling cultivation-based approaches and the uncertain reliability and relative low resolution of in silico predictions - particularly for understudied viral taxa. Towards this, Hi-C proximity ligation uses sequenced, cross-linked virus and host genomic fragments to infer virus-host linkages and has now been applied in at least 10 studies. However, its accuracy remains unknown. Here we assess Hi-C performance in recovering virus-host interactions using synthetic communities (SynComs) composed of four marine bacterial strains and nine phages with known interactions and then apply optimized bioinformatic protocols to natural soil samples. In SynComs, standard Hi-C sample preparations and analyses showed poor normalized contact score performance (26% specificity, 100% sensitivity, incorrect matches up to class level) that could be dramatically improved by Z-score filtering (Z ≥ 0.5, 99% specificity), though at reduced sensitivity (62% down from 100%). Detection limits were established as reproducibility was poor below minimal phage abundances of 105 PFU/mL. Applying optimized bioinformatic protocols to natural soil samples, we compared virus-host linkages inferred from proximity-ligated Hi-C sequencing with predictions generated by in silico homology-based and machine learning-based bioinformatic approaches. Prior to Z-score thresholding, agreement was relatively high at the phylum to family levels (72%), but not at the genus (43%) or species (15%) levels. Z-score thresholding reduced sensitivity (only 34% of predictions were retained), with only modest improvements in congruence with bioinformatic methods (48% or 18% at genus or species levels, respectively). Regardless, this led to 79 genus-level-congruent virus-host linkages and 293 new ones revealed by Hi-C alone, i.e., providing many new virus-host interactions to explore in already well-studied climate-critical soils. Overall, these findings provide empirical benchmarks and methodological guidelines to improve the accuracy and reliability of Hi-C for virus-host linkage studies in complex microbial communities.

Benchmarking↗
PubMed1987

Preclinical approaches to the treatment of metastatic disease: therapeutic properties of rH TNF, rM IFN-gamma, and rH IL-2.

The present studies were undertaken to examine the immunotherapeutic properties of recombinant murine interferon-gamma (rM IFN-gamma), recombinant human tumour necrosis factor (rH TNF), and recombinant human interleukin-2 (rH IL-2) in preclinical metastasis models. It was observed that these cytokines have disparate mechanisms of therapeutic activity as well as different optimal therapeutic protocols. Thus, not only is the dose important to the therapeutic activity of each of the agents; so also is the route of administration, schedule of administration, duration of administration, and sequence of administration. The rM IFN-gamma has a narrow window of activity, with a bell-shaped therapeutic response with a dosage optimum at 50,000 U/animal of rM IFN-gamma administered 3 times per week. In contrast, rH IL-2 has optimal therapeutic activity for the treatment of metastatic disease after i.p. as compared to i.v. administration. This appears to be associated with the serum pharmacokinetics, since longer serum concentrations are achieved following i.p. administration although lower serum levels are also achieved. RH IL-2 has a biphasic dose-response curve for therapeutic activity with optima from 100 to 1000 U/animal and at doses greater than 100,000 U/animal. The lower doses appear to be associated with T cell augmentation whereas the higher doses are associated with NK cell or LAK cell augmentation. RH TNF has therapeutic activity for the treatment of metastatic disease after i.v. but not i.p. administration. High levels of rH TNF are readily detected in the serum following i.v. administration, with a serum half-life of approximately 30 min. In contrast, only minimal serum TNF activity is observed after i.p. administration, suggesting that this may be the origin of the increased therapeutic activity following i.v. administration. Furthermore, rH TNF has additive therapeutic activity when administered in conjunction with suboptimal doses of rM IFN-gamma. Unfortunately, the additive therapeutic activity of rM IFN-gamma and rH TNF is also associated with increased toxicity. However, in preliminary experiments it was found that the b.i.d. administration of aspirin at 25 mg/kg resulted in decreased toxicity. In summary, the recombinant cytokines provide a challenge both preclinically and clinically to the development of optimal therapeutic protocols, and suggest that close attention must be paid to the dose, route, schedule, duration, and sequence of their administration.

Animals↗
PubMed1997

Optimized differential display and reamplification parameters for silver staining.

Differential display (DD) is a powerful instrument to detect differences in gene expression of malignant and normal cells. An optimized silver staining protocol was used to compare mRNA expression of keratinocytes and squamous cell carcinoma cells of the head and neck. Optimal concentration for upstream and downstream primer was 0.2 microM. Best primer concentrations for reamplification were between 40 and 60 pM. With this optimized protocol nearly 50 differentially expressed transcripts have been identified and differential display can be applied safer, easier, and faster, than by radioactive labeling.

Carcinoma, Squamous Cell↗
PubMed1999

Gadolinium contrast-enhanced three-dimensional MRA of peripheral arteries with multiple bolus injection: scan optimization in vitro and in vivo.

In this study, a scanning protocol was developed to image the arterial bed of the pelvis and both legs along their entire length in patients with peripheral arterial disease, using standard hard- and software. Three adjacent stations are acquired consecutively, with some small overlap; per station; one Gadolinium contrast bolus is administered. The scanning protocol was optimized in an in vitro phantom study. The optimal flip angle was found to be 50 degrees. Also, the optimal scan delay was chosen to be equal to the arrival time of the contrast bolus thereby minimizing artifacts. Three contrast bolus injections showed sufficient enhancement of the vessels after image subtraction. Finally, stenosis quantification by manual caliper was performed by five observers in the MRA images and correlated with the percent diameter reduction determined by quantitative angiography from corresponding X-ray images. The results of the MRA measurements were reproducible and intra- and inter-observer variabilities were statistically non-significant (p = 0.54 and p = 0.12, respectively). Stenosis quantification performed by four observers showed a good correlation with the X-ray derived values (rp > 0.90, p < 0.02); the results from one observer were not significantly correlated. Five patients with proven peripheral disease were investigated with this new MRA scanning protocol. The images were of good quality which allowed adequate clinical evaluation; the original diagnoses obtained from X-ray examinations, were confirmed with MRA. In conclusion, peripheral arterial disease can be evaluated adequately with this MR scanning protocol.

Adult↗
PubMedDate not supplied

Determination of the osmotic characteristics of hamster pancreatic islets and isolated pancreatic islet cells.

The ability to store pancreatic islets using cryopreservation methodology would greatly assist the application of clinical islet transplantation to Type 1 (insulin-dependent) diabetics. It is our working thesis that the illumination of fundamental biophysical characteristics of these cells will lead to increased cryosurvival rates through theoretically predicted and experimental testing of optimal freezing protocol; as has been found for cells and tissues such as mammalian and Drosophila embryos. Pancreatic islets were isolated from Golden hamsters and their osmometric behavior, including inactive cell volume (Vb), was determined for either whole islets or isolated individual islet cells. When islets or islet cells were exposed to various concentrations of NaCl, they were found to exhibit a "classic" "Boyle-Van't Hoff" osmometric response. The Boyle-Van't Hoff representation of the volume curve (relative cell volume vs. 1/osmolality) yields a linear response with r values of .99 for each curve. Extrapolations to the normalized osmotically inactive volumes (Vb) were .43 and .22 for whole islets and individual islet cells, respectively. These data regarding the fundamental cryobiological characteristics of islets and islet cells should provide the foundation upon which to further the investigation of osmotic parameters of these cells and eventually lead to the determination of optimal freezing protocols.

Animals↗
PubMed2002

MRI protocol technique in the optimal therapeutic strategy of non-functioning pituitary adenomas.

OBJECTIVES AND DESIGN: We performed a prospective study using magnetic resonance imaging (MRI) at regular post-operative intervals in non-irradiated patients with non-functioning pituitary adenomas (NFAs) to assess the frequency of tumoral regrowth and recurrences, in order to define the indications of post-operative radiotherapy. PATIENTS AND METHODS: Fifty-one patients aged 25--80 years (mean, 55.6plus minus12.3 years) were included. Post-operative MRIs were performed 3--12 months (mean, 5.2plus minus1.7 months) after surgery, 6 months later and then, every 12--18 months for at least 2 years. The mean post-operative follow-up was 67.7plus minus31.8 months (range, 24--144 months). RESULTS: In 17 patients (33%, group I) no tumoral residue was observed on post-operative MRIs and no tumoral recurrence was diagnosed. Tumour regrowth was detected in 13 of the 34 patients (38.2%) with post-operative tumoral residue (group II), 7--66 months (mean, 27.3plus minus17.3 months) after surgery. In this group, Kaplan--Meier analysis showed 78.8% recurrence free survival at 2 years and 60.9% at 5 years. Patients with tumoral regrowth had higher mean residual tumoral volume than patients without any tumoral regrowth in the group II (258plus minus165 vs 163plus minus165 mm(3), P=0.05). CONCLUSIONS: We suggest a MRI protocol that includes, a 4- to 6-, 12- and 24-month post-operative MRI for every patient. When no tumoral residue is seen, pituitary radiotherapy is useless. MRI must be repeated 3, 5 and 10 years after surgery to eliminate late recurrence. The observed frequency of tumoral regrowth in patients with tumoral residue does not justify systematic post-operative radiotherapy. It should be performed only when tumoral regrowth is proved by a yearly MRI survey.

Adenoma↗