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Humans expend energy at rest (REE), and this major energy exchange component is now usually estimated using statistical equations that include weight and other predictor variables. While these formulas are useful in evaluating an individual's or group's REE, an important gap remains: available statistical models are inadequate for explaining underlying organ-specific and tissue-specific mechanisms accounting for resting heat production. The lack of such systems level REE prediction models leaves many research questions unanswered. A potential approach that can fill this gap began with investigators who first showed in animals and later in humans that REE reflects the summated heat production rates of individual organs and tissues. Today, using advanced imaging technologies, REE can be accurately estimated from the measured in vivo mass of 10 organ-tissue mass components combined with their respective mass-specific metabolic rates. This review examines the next frontier of energy expenditure models and discusses how organ-tissue models have the potential not only to better predict REE but also to provide insights into how perturbations in organ mass lead to structure-function changes across other interacting organ systems. The introductory ideas advanced in this review provide a framework for future human energy expenditure modelling research.

The purpose of this study was to model data from a head to head comparison of the in vivo fate of hyper-branched PAMAM dendrimers with linear HPMA copolymers in order to understand the influence of molecular weight (MW), hydrodynamic size (Rh) and polymer architecture on biodistribution in tumor-bearing mice using compartmental pharmacokinetic analysis. Plasma concentration data was modeled by two-compartment analysis using Winnonlin® to obtain elimination clearance (E.CL) and plasma exposure (AUCplasma). Renal clearance (CLR) was calculated from urine data collected over 1 week. A plasma-tumor link model was fitted to experimental plasma and tumor data by varying the tumor extravasation (K4, K6) and elimination (K5) rate constants using multivariable constrained optimization solver in Matlab®. Tumor exposures (AUCtumor) were computed from area under the tumor concentration time profile curve by the linear trapezoidal method. Along with MW and Rh, polymer architecture was critical in affecting the blood and tumor pharmacokinetics of the PAMAM-OH dendrimers and HPMA copolymers. Elimination clearance decreased more rapidly with increase in hydrodynamic size for PAMAM-OH dendrimers as compared to HPMA copolymers. HPMA copolymers were eliminated renally to a higher extent than PAMAM-OH dendrimers. These results are suggestive of a difference in extravasation of polymers of varying architecture through the glomerular basement membrane. While the linear HPMA copolymers can potentially reptate through a pore smaller in size than their hydrodynamic radii in a random coil conformation, PAMAM dendrimers have to deform in order to permeate across the pores. With increase in molecular weight or generation, the deforming capacity of PAMAM-OH dendrimers is known to decrease, making it harder for higher generation PAMAM-OH dendrimers to sieve through the glomerulus as compared to HPMA copolymers of comparable molecular weights. PAMAM-OH dendrimer had greater tumor extravsation rate constants and higher tumor to plasma exposure ratios than HPMA copolymers of comparable molecular weights which indicated that in the size range studied, when in circulation, PAMAM-OH dendrimers had a higher affinity to accumulate in the tumor than the HPMA copolymers.

The effects of floor space on the trailer and journey time during transport from the farm to the packing plant on indicators of stress (open-mouth breathing, muscle tremors, and skin discoloration) and on the incidence of transport losses (dead on arrival, nonambulatory, noninjured, and nonambulatory, injured) were evaluated in a study involving 160 loads of market-weight pigs (BW 124.7 ± 4.38 kg) using a split-plot design with a 2 × 6 factorial arrangement of treatments: 1) journey time [main plot; short ( 0.05) on the incidence of dead and nonambulatory, injured pigs, or on total transport losses. There were interactions (P 0.05) of journey time. The incidence of open-mouth breathing for the 3 smallest floor spaces was greater (P 0.05) of journey time for the 3 greatest floor spaces. The frequency of skin discoloration was greater (P < 0.001) for pigs transported at the 2 smallest floor spaces compared with the other 4 floor spaces. In summary, short journey time increased the frequency of indicators of stress after unloading at the plant for pigs transported at smaller floor spaces and also increased the incidence of nonambulatory, noninjured pigs at 2 of the 3 smallest floor spaces. However, neither transport floor space nor journey time had an effect on total losses.

Spherical and rod-shaped gold nanoparticles with surface poly (ethylene glycol) (PEG) chains were characterized for size, shape, charge, poly dispersity and surface plasmon resonance. The nanoparticles were injected intravenously to 6–8 weeks old female nu/nu mice bearing orthotopic ovarian tumors and their biodistribution in vital organs was compared. Gold nanorods were taken up to a lesser extent by the liver, had longer circulation time in the blood, and higher accumulation in the tumors, compared with their spherical counterparts. The cellular uptake of PEGylated gold nanoparticles by a murine macrophage-like cell line as a function of geometry was examined. Compared to nanospheres, PEGylated gold nanorods were taken up to a lesser extent by macrophages. These studies point to the importance of gold nanoparticle geometry and surface properties on transport across biological barriers.

Body mass index (BMI) is now the most widely used measure of adiposity on a global scale. Nevertheless, intense discussion centers on the appropriateness of BMI as a phenotypic marker of adiposity across populations differing in race and ethnicity. BMI-adiposity relations appear to vary significantly across race/ethnic groups, but a collective critical analysis of these effects establishing their magnitude and underlying body shape/composition basis is lacking. Accordingly, we systematically review the magnitude of these race-ethnic differences across non-Hispanic (NH) white, NH black and Mexican American adults, their anatomic body composition basis and potential biologically linked mechanisms, using both earlier publications and new analyses from the US National Health and Nutrition Examination Survey. Our collective observations provide a new framework for critically evaluating the quantitative relations between BMI and adiposity across groups differing in race and ethnicity; reveal new insights into BMI as a measure of adiposity across the adult age-span; identify knowledge gaps that can form the basis of future research and create a quantitative foundation for developing BMI-related public health recommendations.

The impact of feeding ractopamine hydrochloride (RAC) on growth performance and responses to handling and transport in heavy BW pigs was evaluated in a study performed as a split-plot design with a 3 × 3 factorial arrangement of treatments: 1) RAC level (0 vs. 5 vs. 7.5 mg/kg of feed) and 2) handling intensity (HI; gentle vs. moderate vs. aggressive); RAC level was the main plot and HI was the subplot. A total of 288 pigs housed in groups of 8 were used to evaluate growth performance over a 28-d RAC feeding period (98.5 ± 4.58 to 131.5 ± 7.45 kg BW). On d 29 of the study, the HI treatment was applied to 216 pigs (6/pen; 2/pen on each HI). This was followed by transportation for 1 h on a livestock trailer at the end of which pigs were subjected to a final handling procedure. Blood samples (to measure acid-base, cortisol, and catecholamine levels) were collected and rectal temperature was measured 2 h before the HI treatment (baseline) and after the final handling procedure (final). Feeding RAC (5 and 7.5 mg/kg) improved (0.01) ADG (9.9 and 9.0% for 5 and 7.5 mg/kg RAC, respectively) and G:F (8.8 and 11.8%, respectively) compared to controls, with no differences (0.05) between the 2 RAC levels. Increasing the intensity of handling decreased (0.001) final blood pH, bicarbonate, and base excess and increased (0.001) final blood lactate and plasma cortisol and norepinephrine levels. Aggressive compared to gentle handling increased (0.05) the incidence of pigs exhibiting open-mouth breathing and skin discoloration after the final handling procedure but had no effect (0.05) on the incidence on nonambulatory, noninjured pigs. There was no effect (0.05) of feeding RAC on final rectal temperature or blood acid-base measurements. Feeding 7.5, but not 5, compared to 0 mg/kg RAC increased (0.05) final plasma epinephrine levels and the incidence of nonambulatory, noninjured pigs. This study confirms the improved growth performance of pigs fed RAC and the negative effects of aggressive handling on physical, metabolic, and physiological responses of pigs. It also suggests that pigs fed 5 compared to 0 mg/kg RAC showed similar responses to transport and handling. However, pigs fed 7.5 mg/kg of RAC had a greater incidence of nonambulatory, noninjured pigs when subjected to the handling/transport model and this warrants further investigation.

The ability to achieve high-yield focal plane arrays from Hg1−xCdxTe molecular beam epitaxy material depends strongly on postgrowth wafer analysis. Nondestructive analysis that can determine layer thicknesses as well as alloy compositions is critical in providing run-to-run consistency. In this paper, we incorporate the use of a thin film transmission matrix model to analyze Fourier transform infrared (FTIR) transmission spectra. Our model uses a genetic algorithm along with a multidimensional, nonlinear minimization Nelder-Mead algorithm to determine the composition and thickness of each layer in the measured epitaxial structure. Once a solution has been found, the software is able to predict detector performance such as quantum efficiency and spectral response. We have verified our model by comparing detector spectral data to our predicted spectral data derived from the room-temperature FTIR transmission data. Furthermore, the model can be used to generate design curves for detectors with varying absorber thicknesses and/or different operating temperatures. The consequence of this are reduced cycle times and reduced design variations.

Some men have elevated levels of sperm chromosome aneuploidy. In this study, we have evaluated and summarized sperm aneuploidy rates in male infertility patients and control groups. The mean aneuploidy rate for five chromosomes (X, Y, 13, 18, 21) was 1.2 +/- 0.1 for fertile controls, 1.4 +/- 0.1 for a general population control group, and 5.8 +/- 1.14 for the patients. When the patients were classified by the type of male factor infertility, the total aneuploidy rate was 2.6 +/- 0.3 in men with moderately diminished semen quality (n = 7), 4.0 +/- 0.3 patients with severe teratoasthenooligozoospermia, and 15.9 +/- 3.8 for men with rare ultrastructure defects such as round head only syndrome or severe tail agenesis. Some infertility patients have a severely elevated level of sperm chromosome aneuploidy, which may contribute to infertility or diminish the likelihood of a successful outcome from IVF/ICSI. The severity of sperm chromosome aneuploidy appears to be proportional to the severity of abnormal semen quality: in particular, abnormal morphology. The high rates of aneuploidy in patients with severe ultrastructure defects suggest that caution should be employed in counseling those patients prior to IVF/ICSI.

Purpose:This study was undertaken to evaluate simplified methods of human embryo coculture using either attached or nonattached autologous cumulus tissue. Methods:Eight hundred one zygotes were cultured for 48 hr in a prospective, randomized trial comparing culture of embryos either with intact cumulus tissue, with cumulus tissue added to the droplet of culture medium, or without any cumulus tissue. In a follow-up study, embryo quality, pregnancy rates, and implantation rates were compared in 120 consecutive patients undergoing in vitro fertilization with a coculture system using cumulus tissue compared to a cohort of 127 patients undergoing IVF immediately preceding the institution of the coculture protocol. Results:Embryo morphology was significantly improved (P < 0.05) following culture with attached cumulus tissue (5.61 ± 0.29) and culture with added cumulus tissue (4.72 ± 0.31) compared to that of embryos grown in culture medium without cumulus tissue (3.95 ± 0.26). The clinical pregnancy rate improved from 39.4% (50/127) to 49.2% (59/120) following institution of a system of coculture with attached cumulus tissue. Conclusions:These data indicate that a simple coculture system using autologous cumulus tissue can result in improved embryo morphology, implantation rates, and clinical pregnancy rates during in vitro fertilization. This coculture system is simple, is non-labor intensive, and eliminates many of the risks which may be present in other embryo coculture systems.

Since the first in vitro fertilisation (IVF) pregnancy was delivered in 1978, this procedure has resulted in thousands of pregnancies and opened a vast new frontier of research and treatment for the infertile couple. Pregnancy rates with IVF improve as the number of high quality embryos available for transfer increases; therefore, ovarian stimulation agents to produce multiple oocysts for IVF are advantageous. Clomifene (clomiphene citrate), human menopausal gonadotrophin (hMG; menotropins), and subsequent generations of products are commonly used as stimulation agents. In conjunction with the stimulation agents, gonadotrophin-releasing hormone (GnRH) agonists and human chorionic gonadotrophin (hCG) serve as adjuvants for successful control of all events in the induction process. Clomifene, an estrogen agonists/antagonist, occupies the estrogen receptor for a longer period of time than estrogen (weeks versus hours). Because this signal is interpreted as low estrogen, GnRH is released, which produces a rise in circulating levels of follicle-stimulating hormone (FSH) and luteinising hormone (LH) and subsequent ovarian follicular development. Menotropins is collected by passing urine from menopausal donors over a Sepharose column, followed by removal of high molecular weight impurities by chromatography. The mixture of FSH and LH is biologically standardised. This product stimulates multiple ovarian follicular development. Urofollitrophin is produced using antibodies to hCG anchored to a separation column. LH then can be excluded from the eluate by binding to the hCG antibodies (LH immunoaffinity column). Highly purified FSH is obtained by passing menopausal urine over a column with monoclonal antibodies to FSH. The isolated FSH is then eluted from the column by a highly basic solution and crystallised. This product delivers FSH at a 90% purity and can be administered subcutaneously rather than intramuscularly. Dosage is standardised on a mg/kg basis. Recombinant human FSH is completely free of LH and offers the advantages of better batch consistency, greater purity, and absence of any human contaminants. It may be given both subcutaneously and intravenously. Genetically engineered FSH combines portions of the native protein with another protein (hCG) which enhances its potency and extends the half-life compared with wild-type FSH. Short, medium and ultra-long activity analogues of genetically engineered FSH may be used to tailor stimulation protocols in various clinical situations. Growth hormone is an adjuvant to ovarian stimulation which results in a decreased number of ampoules of menotropins being required to achieve ovulation in poor responders. Ovulation triggers include both hCG and GnRH agonists. Progesterone supplementation is generally used in the luteal phase of the IVF cycle and is administered by intramuscular injection or vaginal suppository. It appears that conscious sedation with midazolam, pethidine (meperidine) and fentanyl is nontoxic for oocyte recovery. If full anaesthesia is required for gamete intrafallopian tube transfer (GIFT) or zygote intrafallopian tube transfer (ZIFT), balanced anaesthesia with nitrous oxide and an opioid appears to be the most appealing option. Appropriate information on the clinical use of the drugs used in IVF greatly reduces patient stress associated with the complex multidrug regimens associated with the procedure.

There is an increased requirement for nutrients in normal pregnancy, not only due to increased demand, but also increased loss. There is also an increased insulin-resistant state during pregnancy mediated by the placental anti-insulin hormones estrogen, progesterone, human somatomammotropin; the pituitary hormone prolactin; and the adrenal hormone, cortisol. If the maternal pancreas cannot increase production of insulin of sustain normoglycemia despite these anti-insulin hormones, gestational diabetes occurs. Gestational diabetes is associated with excessive nutrient losses due to glycosuria. Specific nutrient deficiencies of chromium, magnesium, potassium and pyridoxine may potentiate the tendency towards hyperglycemia in gestational diabetic women because each of these four deficiencies causes impairment of pancreatic insulin production. This review describes the pathophysiology of the hyperglycemia and the nutrient loss in gestational diabetes and further postulates the mechanism whereby vitamin/mineral supplementation may be useful to prevent or ameliorate pregnancy-related glucose intolerance.

The optimum diet composition for weight loss in obese women with or without previous gestational diabetes mellitus remains to be determined. Weight loss may be especially important for the latter group in terms of preventing future gestational as well as non-insulin dependent diabetes mellitus. We studied 40% vs. 55% carbohydrate calorically restricted diets to compare weight loss and metabolic response.We performed a prospective, 12-week, blinded, randomized crossover study of 25 obese women; 13 of whom had previous gestational diabetes. Each woman was allocated to a treatment regimen for 6 weeks and then "crossed over" to the alternative regimen for an additional 6 weeks. Calories were provided in the form of nutritional supplement bars except for the evening meal that comprised 1/3 of the caloric needs. All subjects were seen and weighed weekly. Metabolic variables including glucose tolerance, glycated proteins, lipids, and percent body fat were measured at the beginning, crossover, and end of the study.Women with previous gestational diabetes mellitus were comparable to obese women without a history of previous gestational diabetes except that the former had higher maximum levels of glucose on a glucose tolerance test and higher fasting insulin levels consistent with greater insulin resistance. Weight loss was comparable for all groups during the first 6 weeks but attenuated in all groups during the second 6 weeks of the trial regardless of diabetes history or treatment group allocation. Women with or without a previous history of gestational diabetes had higher triglycerides while on a 55% carbohydrate diet than while on a 40% carbohydrate diet.A weight loss regimen consisting of 40% carbohydrate results in lower triglyceride levels than those achieved with a 55% carbohydrate content diet in obese women. Thus, the hypocaloric diet with the higher fat content produced the more favorable lipid profile in all obese women.

Porcine relaxin has been reported to stimulate various human sperm functions. In this paper we report that human recombinant relaxin binds to human sperm with a high affinity (Kd = 6.5 x 10(-10)). The bound 125I-relaxin was not displaced by insulin, or human chorionic gonadotropin, however, it was displaced by unlabeled relaxin. In sperm function studies, recombinant human relaxin stimulated sperm motility, zona-free hamster egg penetration, and the acrosome reaction.

The optimal availability of immune cells in the peripheral blood streams of women may play a critical role in their response to disease and therapeutic interventions. This study was designed to examine concentrations of circulating white blood cells (WBC) including lymphocyte subsets, during the 24-hr daily and 28-day menstrual cycles. Venous blood (20 ml) from five healthy young women was obtained at 0, 6, 12, and 18 hr on the 6th and 22nd day of the normal menstrual cycle. Cortisol, progesterone (P4), estradiol (E2), total WBC, granulocyte, monocyte, and total lymphocyte levels were measured. Using fluorescent activated cell scanning, levels of T, B, Helper (H), Suppressor (S) and Natural Killer (NK) cells were also determined. Significant differences in the diurnal and Day 6 and 22 means were identified using analysis of variance and the Student's t test. Mean WBC counts differed significantly between individuals and ranged from 3.63 +/- 0.33 to 8.60 +/- 1.00 on Day 6 and 3.75 +/- 0.56 to 9.45 +/- 0.98 on Day 22 (P < 0.05). Fluctuations in the concentrations of peripheral blood immune cells followed a similar pattern for the time points selected. They were lowest in the morning at 6 hr and reached peak concentrations in the evening at 17 hr or at midnight at 24:00 hr. Total WBC and granulocyte levels were consistently highest in the evening at 18 hr while lymphocyte levels either peaked in the evening or at midnight. Between midnight and early morning, levels of WBC, lymphocytes, T, B, H, and S cells all decreased significantly (P < 0.01) and subsequently increased significantly, between 6 hr and noon or noon and 18 hr (P < 0.01). When compared every 12 hr, the levels of WBC, granulocytes, lymphocytes, T, B, H, and S cells showed a significant day time rise between 6 and 18 hr (P < 0.02). NK cells revealed no significant fluctuations for any of the diurnal time point comparisons examined in this study. The WBC means for all subjects on Day 6 was 6.15 +/- 1.96 and on Day 22 was 6.39 +/- 2.14, evidence that the total number of white blood cells was not significantly altered between the 2 days. However when comparing specific time points during the day for the follicular (Day 6) and luteal (Day 22) phases of the menstrual cycle, significant differences were found. Most striking were the monocyte patterns, which revealed a nadir at 12:00 noon on Day 6 and a peak at the same time on Day 22.(ABSTRACT TRUNCATED AT 400 WORDS)

The effects of floor space on the trailer and journey time during transport from the farm to the packing plant on indicators of stress (open-mouth breathing, muscle tremors, and skin discoloration) and on the incidence of transport losses (dead on arrival, nonambulatory, noninjured, and nonambulatory, injured) were evaluated in a study involving 160 loads of market-weight pigs (BW 124.7 ± 4.38 kg) using a split-plot design with a 2 × 6 factorial arrangement of treatments: 1) journey time [main plot; short (1 h) and long (3 h)] and 2) floor space (subplot; 0.396, 0.415, 0.437, 0.462, 0.489, and 0.520 m(2)/pig, which is equivalent to 0.317, 0.332, 0.350, 0.370, 0.391, and 0.416 m(2)/100 kg of BW, respectively). Two consecutively loaded trailers were randomly allotted to journey time treatment. Floor space treatments were compared in the front 3 compartments on the top and bottom decks of the trailer and were created by varying the number of pigs per compartment, which confounds the effect of floor space with group size. Of the 17,652 pigs transported in 954 test compartments, 0.24% died or became nonambulatory. Neither journey time nor floor space had an effect (P0.05) on the incidence of dead and nonambulatory, injured pigs, or on total transport losses. There were interactions (P0.05) between journey time and floor space treatments for the incidences of nonambulatory, noninjured pigs and open-mouth breathing. For 2 of the smallest floor spaces (0.415 and 0.437 m(2)/pig), the incidence of nonambulatory, noninjured pigs was greater on short than on long journeys; for the other 4 floor spaces there was no effect (P0.05) of journey time. The incidence of open-mouth breathing for the 3 smallest floor spaces was greater (P0.05) for short than long journeys, whereas there was no effect (P0.05) of journey time for the 3 greatest floor spaces. The frequency of skin discoloration was greater (P0.001) for pigs transported at the 2 smallest floor spaces compared with the other 4 floor spaces. In summary, short journey time increased the frequency of indicators of stress after unloading at the plant for pigs transported at smaller floor spaces and also increased the incidence of nonambulatory, noninjured pigs at 2 of the 3 smallest floor spaces. However, neither transport floor space nor journey time had an effect on total losses.

Tap-1 and Tap-2 genes code for a heterodimeric peptide transporter required for the normal maturation and surface expression of class I molecules. Polymorphic variants of these MHC encoded genes occur in rats and humans. After failing to amplify a 3' polymerase chain reaction (PCR) product from thymic and splenic cDNA of the nonobese nondiabetic (NON) strain, we considered it possible that Tap-1 polymorphism was present, since cDNA from CBA/J, C57BL/6, BALB/c, and NOD (nonobese diabetic) mice all yielded Tap-1 3' products. Overlapping PCR fragments spanning the highly conserved ATP-binding cassette (ABC) were generated for purposes of restriction endonuclease analysis, studies of IFN-gamma regulation, and sequencing. To avoid amplifying other members of the transporter family, we used a gel-purified 1670-bp Tap-1 PCR "long product" as template for nested PCR. Sequencing revealed three polymorphic alleles. The most divergent was for the NON strain and involved two non-conserved amino acid substitutions (Arg-->Cys397 and Leu-->Arg491) and three silent mutations. NON mice show an abnormal pattern of class I (Kb) expression and a sizeable reduction in the percentage of CD8+ cells in the blood and thymus. In F2 segregants, the low CD8 phenotype mapped to the MHC. Tap-1 genes of NON and C57BL/6 mice were equally sensitive to up-regulation by IFN-gamma. We conclude that the mouse Tap-1 transporter gene, like the Tap-2 of the rat and the Tap-1 and Tap-2 of the human, is polymorphic. The extensive variation and specific codon changes of Tap-1 in the NON mouse raise the possibility that this gene is the MHC locus responsible for altering the intrathymic development of CD8+ T cells.

We present prenatal diagnosis and follow-up examination of an individual with a de novo paracentric inversion of the long are of chromosome 14. A literature search documented 19 other cases of paracentric inversion of 14q. The outcome of each of these cases is specified together with that of this current case. Four of the 20 cases, all XY, manifested significant abnormalities with mental retardation and microcephaly present in 3 of the 4 cases; 15% (2/13) of familial cases had abnormalities and 40% (2/5) of de novo cases had abnormalities.

Water deficits during flowering decrease the number of seed-bearing pods in soybean (Glycine max L. Merr.). Failure to set pods may indicate an inherent sensitivity to low tissue water potential (Ψ w ) at critical stages of flower development. To test this, we measured the water status and pod set of flowers exposed to water deficits at various stages of development from bud to early ovary (pod) expansion. In well-watered (control) plants, the Ψ w of buds, flowers, and pods were about 0.2 to 0.3 MPa lower than leaf Ψ w at midday

This study analyzed the role that the cumulus oophorus may play in the selection of morphologically normal sperm and the induction of the acrosome reaction. Using the triple stain technique, sperm morphology and acrosomal status were compared between sperm that penetrated the cumulus during in vitro fertilization and sperm from outside the cumulus. The mean percentage of morphologically normal sperm in the samples was 54 +/- 2.8 and increased (p0.05) to 67 +/- 2.6 within the cumulus. Tapered sperm were significantly decreased (p0.05) within the cumulus. The percentage of sperm undergoing the acrosome reaction significantly increased (p0.05) from 14.5 +/- 1.5 to 24.5 +/- 1.9 when incubated with a cumulus mass, and further increased to 49 +/- 3.3 when incubated with mature, expanded cumulus tissue containing an oocyte. These data indicate that human cumulus oophorus plays an active role in the selection of morphologically normal sperm, and influences the ability of the sperm to undergo the acrosome reaction.

The biodistribution profile of a series of linear N-(2-hydroxylpropyl)methacrylamide (HPMA) copolymers was compared with that of branched poly(amido amine) dendrimers containing surface hydroxyl groups (PAMAM-OH) in orthotopic ovarian-tumor-bearing mice. Below an average molecular weight (MW) of 29 kDa, the HPMA copolymers were smaller than the PAMAM-OH dendrimers of comparable molecular weight. In addition to molecular weight, hydrodynamic size and polymer architecture affected the biodistribution of these constructs. Biodistribution studies were performed by dosing mice with (125)iodine-labeled polymers and collecting all major organ systems, carcass, and excreta at defined time points. Radiolabeled polymers were detected in organ systems by measuring gamma emission of the (125)iodine radiolabel. The hyperbranched PAMAM dendrimer, hydroxyl-terminated, generation 5 (G5.0-OH), was retained in the kidney over 1 week, whereas the linear HPMA copolymer of comparable molecular weight was excreted into the urine and did not show persistent renal accumulation. PAMAM dendrimer, hydroxyl-terminated, generation 6.0 (G6.0-OH), was taken up by the liver to a higher extent, whereas the HPMA copolymer of comparable molecular weight was observed to have a plasma exposure three times that of this dendrimer. Tumor accumulation and plasma exposure were correlated with the hydrodynamic sizes of the polymers. PAMAM dendrimer, hydroxyl-terminated, generation 7.0 (G7.0-OH), showed extended plasma circulation, enhanced tumor accumulation, and prolonged retention with the highest tumor/blood ratio for the polymers under study. Head-to-head comparative study of HPMA copolymers and PAMAM dendrimers can guide the rational design and development of carriers based on these systems for the delivery of bioactive and imaging agents.

Establishment of dual cultures of slash pine (Pinus elliottii Engelm. var. elliottii) with vegetative propagules of Fusarium moniliforme var. subglutinans (FMS) was only partially successful. In contrast, the response of slash pine callus to FMS culture filtrate was significant. The in vitro selection for FMS resistance using culture filtrate and slash pine callus or plantlets merits further investigation.

Endosperm development in ovules was studied from the time of fertilization through the P5 stage in setting flowers of soybean and provides a basis for comparison with ovules of similar age from flowers programed to abscisse

6-benzylaminopurine (BAP) delays leaf abscission of soybean Glycine max (L.) Merr. Abscission of the distal pulvinus of primary leaves was induced in 12-d-old seedlings or explants by removal of the leaf blade. BAP applied to the cut end of the pulvinus following leaf blade removal delayed abscission. Discoloration of the pulvinus occurred before abscission commenced and the number of grana in chloroplasts within cortical parenchyma cells of the pulvinus decreased over time following leaf blade removal

Flower and pod abscission limit soybean yield. A system for quantifying flower and pod development based on the morphological appearance of the flower prior to and following anthesis has been developed to aid in studies of pod abscission. Changes in the appearance of the corolla, primarily the banner petal, are used to distinguish the different stages of the system. External pistil dimensions have been correlated with internal features for each stage of development. From anthesis to pod set, pistil length and weight increase almost two- and fivefold, respectively, and ovule development progresses from unfertilized egg cells to embryos surrounded by cellular endosperm. Pod dimensions are correlated with ovule length and width and embryo cell number. Flower and pod stages can be determined in situ, thus permitting non-destructive observation and experimental manipulation of flowers or pods without necessarily impeding their development. Stages have been identified that indicate precisely when pod set occurs and when young pods cease growing and ultimately abscise. This system of flower and pod staging is useful in studies designed to assess effects of abiotic or biotic stress and genetic factors on pod set and abortion.

The cytokinin 6-benzylaminopurine (BAP) increases pod set of soybean Glycine max (L.) Merr. This study was performed to determine the effect of site and method of BAP application on pedicel abscission and the accompanying changes in rachis anatomy. Spraying racemes with BAP in solution, or applying in a lanolin suspension to proximal nodes on a rachis where proximal pedicels had been excised, delayed pedicel abscission at distal nodes. Applying BAP in lanolin to distal pedicels following flower excision failed to delay their abscission. BAP caused rachis swelling only when pods were present, but BAP could delay pedicel abscission either in the presence or absence of pods. These results suggest that rachis swelling following BAP treatment does not have a causal relationship to a delay or decrease of pedicel abscission.

Blood samples were obtained from miniature swine maintained on 0, 2, or 6 g/kg/24 hr ethanol for 8 months (N= 6 in each group). Samples from drinking pigs were taken after 8 hr of ethanol abstinence and all were coded and sent for “blinded” analysis. A fluorigenic high performance liquid chromatographic assay was used to quantify whole blood-associated acetaldehyde, hemoglobin-associated acetaldehyde, plasma-associated acetaldehyde, platelet-associated acetaldehyde, and lymphocyte-associated acetaldehyde. Detectable levels of acetaldehyde were found in each sample in both drinking and nondrinking pigs. Analysis of whole blood-associated acetaldehyde was most discriminatory in distinguishing nondrinking from drinking pigs (mean 21.4 ± 1.0 μM for nondrinkers vs. 24.6 ± 1.5 so for the group consuming 2 g/kg ethanol, p= 0.001). Measurements of hemoglobin-associated acetaldehyde normalized to protein concentration (250 ± 47 nmoles/g vs. 203 ± 33 SD, p < 0.05 drinking vs. nondrinking pigs) and platelet-associated acetaldehyde (0.46 0.34 vs. 0.15 ± 0.16 nmoles/3 × 10° platelets, p= 0.05 drinking vs. nondrinking pigs) were also useful in discriminating drinking from nondrinking animals. Analysis of plasma-associated acetaldehyde and lymphocyte-associated acetaldehyde were not useful as markers of ethanol consumption. The miniature swine proved to be a useful model for the evaluation of these potential markers of ethanol intake. In this model, increased amounts of acetaldehyde can be found in several blood compartments eight hr after cessation of ethanol. Further work is warranted to define the relative kinetics of each measurement and the potential utility of each as a marker for ethanol consumption in man.

Lesser cornstalk borer, Elasmopalpus lignosellus (Zeller) (Lepidoptera: Pyralidae), larval injury to the root-hypocotyl region of greenhouse-reared ‘Florunner’ peanut, Arachis hypogaea L., plants was investigated by determining the size and distribution of feeding sites. The percent injury to periderm, cortex, phloem, and xylem tissues also was estimated. Tissue loss due to insect feeding was measured for only the most prominent of several wound sites on each stem axis. Percent injury to principal tissues was estimated by planimetric methods. The lesser cornstalk borer caused significant injury to the periderm, cortex, and the phloem on mainstems of peanut plants. Mean percent injury was 31.67% for the periderm and cortex, 16.78% for phloem, and only 0.19% for the xylem. Approximately 91% of all feeding sites occurred on the mainstem or were contiguous with the mainstem and a branch. The frequently of plants with feeding sites declined linearly with increasing number of sites. These data support the hypothesis that root growth declines with increasing larval injury to phloem and xylem (food and water conducting tissues, respectively) in the root-hypocotyl region.

Previous studies have indicated that sperm quality may be related to unexplained recurrent pregnancy loss. This study evaluated the degree of sperm DNA fragmentation using the TUNEL assay on sperm from 24 couples with unexplained recurrent pregnancy loss (RPL) compared to sperm from 2 control groups: donors of known fertility and unscreened men from the general population. The percentage of sperm staining positive for DNA fragmentation was increased (p < .001) in the RPL group (38 +/- 4.2) compared to the donor (11.9 +/- 1.0) or general population (22 +/- 2.0) control groups. In the RPL group, no correlation was observed between semen quality parameters and the TUNEL data. These data indicate that some RPL patients have a significant increase of sperm DNA fragmentation, which may be causative of pregnancy loss in some patients.

Background Catamenial pneumothorax, a rare complication of systemic endometriosis, has been difficult to treat successfully. Successful medical therapy is associated with amenorrhea. Case A 44-year-old white woman with recurring catamenial pneumothorax underwent thoracotomy and abrasive pleurodesis. Following the procedures, pneumothorax occurred again and she was treated with the GnRH analogue leuprolide acetate, 3.75 mg monthly intramuscularly. After 6 months, her therapy was changed to continuous hormonal suppression with norethindrone, 0.7 mg/day. After 6 months of this therapy and into the third episode of vaginal bleeding, the patient had another recurrent pneumothorax. Conclusion Leuprolide acetate followed by continuous hormonal suppression with norethindrone was successful for 1 year in resolving recurring postsurgical catamenial pneumothorax, but the problem recurred with the resumption of vaginal bleeding during progestin therapy. Successful medical therapy requires amenorrhea.

This article summarizes the proceedings of a workshop presented at the 2000 RSA Meeting in Denver, Colorado. The aim of this workshop was to discuss the basic methodologies, diagnostic performance, and clinical utility of three technologies: carbohydrate-deficient transferrin, the "Early Detection of Alcohol Consumption" score, and whole blood associated acetaldehyde. Each method adopts a different strategy to identify heavy alcohol consumption and offers a unique approach to determine alcohol abstinence and relapses. Appropriate application of these technologies can lead to early intervention for alcohol problems before significant tissue damage occurs. To date these methodologies have yet to be formally contrasted and compared. Sensitivity, specificity, predictive value, availability, ease of use, and interpretation of tests results are important aspects to consider when selecting the most appropriate and cost-effective system. Critical evaluation of these methodologies can enable research and clinical laboratories to choose the system that best meets their particular needs in terms of assay feasibility, budget, and goals.

Fetal pancreatic tissue has been suggested to be less immunogenic than adult islets. Thus, transplantation of human fetal pancreatic tissue as treatment for type 1 diabetes has been gaining interest. To investigate this question, we tested the peripheral blood mononuclear cell (PBMC) responses from different subject populations to human adult islet proteins (AIP) versus human fetal pancreatic proteins (FPP).PBMC responses to FPP and AIP from normal controls (n=14), newly diagnosed type 1 diabetes patients (n=5), long-term type 1 diabetes patients (n=9), and subjects at-risk for development of type 1 diabetes (n=3) were studied.We observed that normal controls demonstrated PBMC reactivity to 0-3 molecular weight regions (mwr) for both the AIP (mean+/-SD, 0.8+/-1.1) and the FPP (0.6+/-0.7). In contrast, newly diagnosed type 1 diabetic patients (1 year) demonstrated PBMC responses to 9-16 mwr for the AIP (12.8+/-2.5) and 0-14 mwr for the FPP (6.8+/-5.0). The PBMCs from long-term type 1 diabetes patients (3 years) were responsive to 2-11 mwr for AIP (6.0+/-2.8) and 0-11 mwr for FPP (4.9+/-4.0). Three nondiabetic ICA positive subjects at-risk for development of type 1 diabetes demonstrated positive PBMC reactivity to 9-18 mwr for the AIP (12.7+/-3.9) and 4-18 mwr for the FPP (10.0+/-5.9).We conclude that human fetal pancreatic proteins are not significantly less stimulatory than human adult islet proteins to PBMCs of subjects with or at risk for type 1 diabetes.

This study demonstrates the selective tumor targeting and the antitumor efficacy of the N-(2-hydroxypropyl)methacrylamide (HPMA) copolymer-bound mesochlorin e6 monoethylenediamine (Mce6) and HPMA copolymer-bound Adriamycin (ADR) in combination photodynamic therapy (PDT) and chemotherapy against human ovarian OVCAR-3 carcinoma xenografted in female athynmic mice. The concentrations of Mce6 and ADR in blood and tissues, in free or HPMA copolymer-bound form, were determined by fluorescence and high-performance liquid chromatography fluorescence assays, respectively. Xenograft responses to single and combination therapies were recorded. The peak concentration of HPMA copolymer-Mce6 conjugate in tumor was achieved 18 h after administration. For HPMA copolymer-bound drugs, the concentration ratios of liver and spleen versus muscle were significantly higher than those of free drugs. The HPMA copolymer-bound drugs demonstrated selective targeting and accumulation in the tumor, probably attributed to the enhanced permeability and retention effect. In vivo studies revealed that all tumors in the treatment groups showed significant responses after receiving any of the various types of therapy as compared with controls (P0.001). PDT with HPMA copolymer-Mce6 conjugate (PDTMC) at a dose of 13.4 mg/kg (1.5 mg/kg of Mce6 equivalent) and light doses of 110 J/cm2 at 12 and 18 h, respectively, resulted in significant suppression of the growth of OVCAR-3 tumors. Three courses of chemotherapy using 35 mg/kg (2.2 mg/kg of ADR equivalent) of HPMA copolymer-ADR conjugate (CHEMO) were effective in suppressing the growth of tumors. Single PDTMC plus multiple CHEMO exhibited significantly greater therapeutic efficacy than multiple CHEMO. In the group of mice receiving multiple PDTMC, tumor recurrence became obvious after day 20. However, 10 of 12 tumors exhibited complete responses in the group of mice receiving multiple PDTMC plus multiple CHEMO. The least to most effective treatments were ranked as follows: multiple CHEMOsingle PDTMC plus multiple CHEMOmultiple PDTMCmultiple PDTMC plus multiple CHEMO. The results clearly demonstrate that: (a) HPMA copolymer-bound drugs exhibited selective tumor accumulation contrary to free drugs; (b) PDT using HPMA copolymer-Mce6 conjugate with multiple light irradiations was a better therapy than that with single light irradiation; and (c) combination chemotherapy and photodynamic therapy with HPMA copolymer-ADR and HPMA copolymer-Mce6 conjugates was the most effective regimen.

Insulin-dependent (type 1) diabetes is a cell-mediated autoimmune disease. Successful transplantation of human fetal pancreatic tissue into type 1 diabetic patients must address both autoimmunity and allograft rejection. We investigated whether humoral and cellular responses to islet antigens could be demonstrated in the peripheral blood of type 1 diabetic subjects receiving human fetal pancreatic tissue transplants.We investigated peripheral blood mononuclear cell (PBMC) responses, using cellular immunoblotting, and autoantibody responses to islet proteins, before transplantation and at 3-month intervals after transplantation. Our study population included nine long-term type 1 diabetes patients (mean disease duration of 21 years) receiving human fetal pancreatic tissue subcutaneously into the abdominal wall without immunosuppression.Before transplantation, all nine subjects tested negative for islet cell autoantibody (ICA), and seven of nine subjects tested positive for glutamic acid decarboxylase autoantibody (GADAb). After transplantation, all subjects became ICA(+), and the two patients who were GADAb(-) before transplantation, became GADAb(+) after transplantation. Maximum PBMC reactivity to separated human fetal pancreatic proteins was observed in four patients at 3 months, in one patient at 6 months, in two patients at 9 months, and in one patient at 12 months after transplantation. One subject, who had PBMC reactivity to multiple islet proteins before transplantation, continued to respond to multiple islet proteins throughout the study.We conclude that the development in the peripheral blood of ICA, GADAb, and PBMC reactivity to human fetal pancreatic proteins in the transplant recipients is most consistent with reactivation of the type 1 diabetes disease process.

Two isoforms of prostaglandin G/H synthase, PGS-1 and PGS-2, catalyze the formation of prostaglandins (PG). Nonselective PGS inhibitors, e.g., indomethacin, reduce the number of ovulations and PG levels in many animal models. This study evaluated the effects of the selective PGS-2 inhibitor NS-398, compared to indomethacin, on ovulation number and on PG and steroid production both in vivo and in vitro in the rat. NS-398 reduced the synthesis of PGE2 in isolated, LH-stimulated preovulatory follicles incubated in vitro. The inhibition by NS-398 was similar to that of indomethacin. Maximal inhibition was noted from 0.1 microM. Neither progesterone nor cAMP production was affected by NS-398 or indomethacin. The effect of in vivo administration of NS-398 (1, 3, or 10 mg/kg BW, s. c.) to proestrous rats 1 h after the injection of an ovulatory dose of hCG was monitored in follicles extirpated 10 h after hCG. These follicles were incubated in vitro, and NS-398 dose-dependently reduced PGE2 production. The synthesis of cAMP and progesterone was not altered. In separate experiments, the same doses of NS-398 were injected to determine their effect on ovulation in vivo. The number of ovulations was decreased by the highest dose of NS-398. In the in vitro ovarian perfusion model, NS-398 (10 microM) reduced the number of ovulations initiated by LH and isobutylmethylxanthine. Lower doses of NS-398 (0.1 and 1 microM) were less effective. The production of prostanoids (PGE2, PGF2alpha, and 6-keto-PGF1alpha) was reduced in a dose-dependent manner by NS-398. The secretion of steroids was not affected. This study demonstrates that selective inhibition of PGS-2 by NS-398 reduces LH/hCG-stimulated production of prostanoids and the number of ovulations both in vivo and in vitro. These results provide direct evidence to strengthen the role of the inducible, granulosa cell-expressed PGS-2 as one of the key regulators in the ovulatory process and also document that the elevated and perhaps sustained levels of PG are obligatory for ovulation.

Hemoglobin A1c (HbA1c) is a stable minor Hb variant formed in vivo by posttranslational modification by glucose, originally identified by using cation exchange chromatography, and containing primarily glycated N-terminal beta-chains. However, the structure(s) of the quantified species has not been elucidated, and the available methods lack a reference standard. We used electrospray ionization mass spectrometry to determine the extent of glycation of samples separated by boronate affinity and/or cation exchange chromatography. Analyses of clinical samples were consistent with the curvilinear relationship of patient glucose and HbA1c. As glycation increased, the ratio of beta-chain to alpha-chain glycation increased, and the number of glycation sites on the beta-chain increased, although these were relatively minor components. We found several glycated species that cochromatographed with HbA1c on cation exchange, including species with both glycated alpha- and beta-chains, nonglycated alpha- and glycated beta-chains, and multiply glycated beta-chains. The combined use of affinity and cation exchange chromatography with structural confirmation by electrospray ionization mass spectrometry was found to be useful in producing samples of sufficient purity for the standardization of glycohemoglobin clinical assays.

Eicosanoids, the active metabolites of arachidonic acid, are grouped into cyclooxygenase products (prostaglandins [PGs] and thromboxanes) and lipoxygenase products (leukotrienes [LTs] and lipoxins). Numerous studies suggest a role for the lipoxygenase system in ovulation. The aim of this study was to further characterize the effects of lipoxygenase inhibition and the interactions of the lipoxygenase and cyclooxygenase systems in the rat ovary during ovulation. The lipoxygenase inhibitor, nordihydroguaiaretic acid (NDGA), was administered in vivo and in the isolated perfused rat ovary to determine its effect on ovulation rate. The in vivo study confirmed the inhibitory effect of NDGA, and in the perfusion experiments, NDGA caused a dose-dependent reduction in the ovulation rate. To further define the interaction between the lipoxygenase and cyclooxygenase systems, a second set of perfusions was performed with NDGA (10 microM) and the combination of NDGA (10 microM) plus a nonselective cyclooxygenase inhibitor, indomethacin (10 microM). NDGA significantly reduced the number of ovulations compared to that in controls. The ovulation rate for the combination of NDGA+indomethacin was also significantly lower than in controls but not different from that in the NDGA-treated group. Steroidogenesis was decreased only in the NDGA+indomethacin perfusions. Ovarian tissue PGE2 and PGF2alpha levels in the NDGA-treated ovaries were significantly suppressed compared to those in controls. Almost a complete block of PGE2 and PGF2alpha was seen in the NDGA+indomethacin group. LTB4 levels in the 10-h-perfused ovarian tissues were significantly decreased by NDGA compared to those in control tissues. Furthermore, LTB4 (3 microg added twice) completely reversed the inhibitory effect of 0.1 microM NDGA on ovulation rate and partially reversed the effect of 10 microM NDGA in the perfusion model. These results demonstrate that the products of the lipoxygenase pathway, especially LTB4, are important in the process of ovulation in this cyclically ovulating species. The interconnected lipoxygenase and cyclooxygenase pathways may optimize ovulation and facilitate steroidogenesis.

This study characterizes the efficacy and toxicity of: (a) free Adriamycin and N-(2-hydroxypropyl) methacrylamide (HPMA) copolymer-Adriamycin conjugate (P-A); (b) free and HPMA copolymer-meso-chlorin e6 monoethylene diamine disodium salt (Mce6) conjugate (P-C) and light-induced photodynamic therapy; and (c) combinations of the HPMA copolymer conjugates (P-A and P-C) in the destruction of human epithelial ovarian carcinoma heterotransplanted in the nude mouse (OVCAR-3). Eight-week-old female nu/nu mice were injected in both flanks with 0.04-0.05 cm3 OVCAR-3 solid tumor dispersed in media. When bilateral tumors reached a minimum volume of 0.18 cm3 (one axis, 2.0-mm minimum) and demonstrated consistent growth, the experiments were initiated. Drugs were given i.v. unless otherwise noted. Tumor-bearing mice were allocated to the following protocols: (a) Adriamycin at 1 mg/kg, P-A at 30 mg/kg (2.2 mg/kg Adriamycin equivalent), and controls (n = 6 each); (b) Mce6 and light (2 h after administration: 650 nm light for 15 min to deliver 220 J/cm2) at 1.25, 2.5, 5, and 10 mg/kg (n = 6 each), 2.5 mg/kg i.p. (n = 4), and controls (n = 6); (c) P-C at 12.5, 25, and 75 mg/kg (1.5, 2.9, and 8.7 mg/kg Mce6 equivalent, respectively with light (18 h after administration; 650 nm light for 15 min to deliver 220 J/cm2), P-C at 25 mg/kg (2.9 mg/kg Mce6 equivalent) with no light administration, and controls (n = 7 each); and (d) a combination of P-A (30 mg/kg, 2.2 mg/kg adriamycin equivalent) and P-C (12.5 and 75 mg/kg, 1.5 mg/kg and 8.7 mg/kg Mce6 equivalent, respectively) with and without light (n = 7 each; 18 h after administration; 650 nm light for 15 min to deliver 220 J/cm2) and controls (n = 12). Tumor volumes and animals weights were assessed for significant differences from the treated and controls groups by Student's t test. Adriamycin (1 mg/kg) and P-A (30 mg/kg. 2.2 mg/kg Adriamycin equivalent) caused less than a 10% weight loss, and treated tumor volumes (day 10-32) were significantly less than those of controls (all P0.045). Mce6 (2.5-10 mg/kg i.v.), caused tumor regression in 80% of tumors and a shock syndrome in 17-83%. i.p. dosing (2.5 mg/kg) was uniformly fatal. Mce6 at 1.25 mg/kg did not show reproducible efficacy. P-C with light (25 and 75 mg/kg, 2.9 and 8.7 mg/kg Mce6 equivalent, respectively) demonstrated significant tumor destruction (P0.003) but not complete ablation. The combinations of P-A (30 mg/kg, 2.2 mg/kg Adriamycin equivalent) plus P-C (12.5 and 75 mg/kg; 1.5 mg/kg and 8.7 mg/kg of Mce6 equivalent, respectively) with light resulted in tumor volumes that were significantly less than control tumor volumes and the tumor volumes of mice receiving either P-A (30 mg/kg, 2.2 mg/kg Adriamycin equivalent) or P-C with light (12.5 or 75 mg/kg. 1.5 or 8.7 mg/kg Mce6 equivalent, respectively) alone (all P0.02). P-C (75 mg/kg, 8.7 mg/kg Mce6 equivalent) added to P-A (30 mg/kg, 2.2 mg/kg Adriamycin equivalent) resulted in complete tumor ablation. Free Mce6 demonstrates a narrow margin of safety, which is extended by incorporation into HPMA copolymers. P-A demonstrates safety and efficacy in vivo. The combined chemotherapy and photodynamic therapy of P-A (30 mg/kg, 2.2 mg/kg Adriamycin equivalent) with P-C and light (12.5 and 75 mg/kg 1.5 and 8.7 mg/kg Mce6 equivalent, respectively) was nontoxic and allowed us to attain a significant improvement in tumor cures than those obtained by P-A or P-C with light alone.

A polypoid uterine hemangioendothelioma was treated by conservative means.The diagnosis of hemangioendothelioma was confirmed by histopathology and angiography. The patient desired to retain her fertility; therefore, the hemangioendothelioma was treated by hysteroscopically guided Nd:YAG laser ablation.Five years of follow-up by hysteroscopy and dilation and curettage reveal no evidence of recurrence. The patient continues to have regular menses.Conservative management of a uterine hemangioendothelioma by Nd:YAG laser ablation was successful over 5 years of follow-up.

Peritoneal inclusion cysts have not received the attention they merit in the imaging literature. We present a series of peritoneal inclusion cysts and describe their sonographic features. Our findings lead us to encourage more conservative therapies. All seven patients in our series had pelvic pain and had undergone surgery previously. An ovary surrounded by septations and fluid was the most common finding by transvaginal sonography. Doppler examination showed low resistive flow in the septations. Conservative therapy was used in five cases with excellent results. We found that a confident diagnosis of peritoneal inclusion cysts is possible with ultrasonography. This diagnosis should encourage the use of more conservative therapy.

Considering the differing mechanisms of cytotoxicity produced by adriamycin and the photosensitizer meso-chlorin e6 monoethylene diamine (Mce6) with light, the interaction of these agents in combination on human ovarian epithelial carcinoma (OVCAR-3 in vitro) was evaluated by dose and effect addition isobole analysis.Mitochondrial respiration via the 3-(4,5-dimethyl thiazol-2 yl)-2,5-diphenyl tetrazolium bromide cleavage assay (MTT) and reproductive capacity via the tritiated thymidine incorporation assay (TI) were assessed 72 and 144 hours after exposure to adriamycin, Mce6, and light (650 nm), and to their combinations, in OVCAR-3 cells grown in vitro (20,000 cells per well).In the majority of assays, reproductive capacity was more sensitive to the drug(s) than was mitochondrial respiration (2-10x). Dose-addition isobole analysis showed synergy for the combination of 50% median effective dose (ED50) adriamycin with 50% ED50 Mce6/light in all assays (all Por = .027). Antagonism was noted with the combination 25% ED50 adriamycin with 75% ED50 Mce6/light. Additivity and synergy were the predominant interactions for 75% ED50 adriamycin with 25% ED50 Mce6/light by dose-addition isobole analyses. Effect-addition isoboles showed a predominance of synergy, particularly for the combination 50% ED50 adriamycin with 50% ED50 Mce6/light.Synergy and additivity are the primary in vitro interactions for the combination of adriamycin and Mce6/light in the dosage range tested. Reproductive capacity is more sensitive to these agents than is mitochondrial respiration.