Hydraulic platform to permit progressive resisted exercises in a Hubbard tank.
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The obscured pronuclei or nuclei in living one- and two-celled pig ova were revealed after centrifugation for 3 min at 15,000 X g. To determine viability of centrifuged ova, one- and two-celled pig ova were collected from superovulated gilts; half of the ova were centrifuged and all ova were transferred into recipient gilts. Prior to transfer all embryos were stained with tetramethylrhodamine isothiocyanate (TRITC) to distinguish the experimental embryos from the recipients's own ova. Centrifuged ova were transferred into one oviduct of recipient gilts and uncentrifuged ova were deposited into the opposite oviduct. Embryos were recovered 4 days after transfer and were stained with lacmoid or Hoechst 33342 to assess their stage of development. Centrifugation had no detectable influence on survival of the recovered embryos to 4 days. Centrifugation is a simple, reliable method for revealing pronuclei and nuclei of one- and two-celled pig ova and apparently does not alter subsequent preimplantation development.
Exposure of mammalian oocytes to the protein phosphatase (PP)-1 (PP1) and PP2A inhibitor okadaic acid (OA) stimulates oocyte meiosis. However, treated oocytes do not develop beyond metaphase I (MI), and they display morphological aberrations. Experiments were conducted to define inhibitor treatment conditions for macaque oocytes that would result in germinal vesicle breakdown (GVB) stimulation and completion of meiosis without significant cytoplasmic abnormalities. As described above for OA, continual exposure of macaque oocytes to 50 nM calyculin-a (CL-A) significantly enhanced GVB at 24 h compared to that in controls, and the majority of the treated oocytes displayed cytoplasmic abnormalities. However, transient exposure (10 min) of rhesus macaque oocytes to either 50 nM CL-A or 1.0 microM OA enhanced GVB rates compared to that in controls and did not increase the incidence of cytoplasmic abnormalities. Meiotic maturation from germinal vesicle-intact oocytes to MII was enhanced following transient treatment with CL-A or OA compared to that in controls; however, development from MI to MII occurred at a similar frequency. In vitro-matured oocytes transiently exposed to OA and CL-A were capable of fertilization. In addition, ovarian immunohistochemical analysis revealed that both PP1 and PP2A were present in macaque oocytes. PP1 was localized throughout the cytoplasm with a predominance in the nucleus, whereas PP2A was evenly distributed throughout the cytoplasm with a reduction in the nuclear area. These results taken together-differential developmental responses to inhibitor treatment and intracellular enzyme localizations-may be indicative of multiple regulatory roles of PP1 and/or PP2A during meiosis.
Colony-stimulating factors (CSFs) shorten the duration of myelosuppression following chemotherapy and, thus, allow the administration of higher doses. This study evaluates the efficacy of granulocyte macrophage colony-stimulating factor (GM-CSF) and granulocyte colony-stimulating factor (G-CSF) in allowing administration of high-dose cyclophosphamide in combination with doxorubicin. Ninety women with metastatic, locally advanced, or high-risk (> or = 10 positive nodes) breast cancer and no prior anthracycline treatment were given doxorubicin (60 mg/m2) with progressively increased doses of cyclophosphamide (1,200 mg/m2, 1,800 mg/m2, and 2,400 mg/m2). The first 60 patients received GM-CSF; the remaining 30, G-CSF. The maximum tolerated dose was not reached with 2,400 mg/m2 of cyclophosphamide. When compared to GM-CSF, G-CSF significantly reduced the duration of granulocytopenia (P < .001). No differences in duration of thrombocytopenia were noted. The results were not sufficiently consistent to indicate a trend toward reduction in rates of febrile neutropenia with one CSF versus the other. However, patients who received G-CSF were hospitalized less frequently than those receiving GM-CSF. With CSFs, high-dose cyclophosphamide in combination with doxorubicin can be safely administered on an outpatient basis. A shorter duration of granulocytopenia resulted from the use of G-CSF than from GM-CSF.
UNLABELLED: Administrators routinely seek to increase contribution margin (revenue minus variable costs) to better cover fixed costs, provide indigent care, and meet other community service responsibilities. Hospitals with high operating room (OR) utilizations can allocate OR time for elective surgery to surgeons based partly on their contribution margins per hour of OR time. This applies particularly when OR caseload is limited by nursing recruitment. From a hospital's annual accounting data for elective cases, we calculated the following for each surgeon's patients: variable costs for the entire hospitalization or outpatient visit, revenues, hours of OR time, hours of regular ward time, and hours of intensive care unit (ICU) time. The contribution margin per hour of OR time varied more than 1000% among surgeons. Linear programming showed that reallocating OR time among surgeons could increase the overall hospital contribution margin for elective surgery by 7.1%. This was not achieved simply by taking OR time from surgeons with the smallest contribution margins per OR hour and giving it to the surgeons with the largest contribution margins per OR hour because different surgeons used differing amounts of hospital ward and ICU time. We conclude that to achieve substantive improvement in a hospital's perioperative financial performance despite restrictions on available OR, hospital ward, or ICU time, contribution margin per OR hour should be considered (perhaps along with OR utilization) when OR time is allocated. IMPLICATIONS: For hospitals where elective surgery caseload is limited by nursing recruitment, to increase one surgeon's operating room time either another surgeon's time must be decreased, nurses need to be paid a premium for working longer hours, or higher-priced "traveling" nurses can be contracted. Linear programming was performed using Microsoft Excel to estimate the effect of each of these interventions on hospital contribution margin.
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We have developed and evaluated an ELISA-based collagen binding assay (CBA) as an aid in the diagnosis and classification of von Willebrand's disease (vWD). The assay is simple to perform, and appears capable of differentiating Type II vWD from Type I vWD. Using plasma samples from both affected and non-affected patients, or from normal individuals, data obtained using the CBA were directly compared to data simultaneously derived from a standard von Willebrand factor antigen (protein; vWFAg) ELISA, and from a standard ristocetin cofactor (RCof) assay. Plasma derived from vWD patients (both Type I and Type II) showed overall reduced levels of vWF as detected by all three assays. Mean levels as a per cent of normal for vWFAg, CBA, RCof were 47.3, 60.7, 31.1 for Type I patients (n = 37), and 34.9, 1.6, 11.9 for Type II patients (n = 16) respectively. However despite the reduced vWF levels detected in Type I vWF binding values for both the CBA and vWFAg showed near comparability (i.e. vWFAg:CBA ratio generally less than or equal to 1.0). These ratio values were thus similar to those observed using plasma derived from either individual normal donors, or from non-vWD affected patients. On the other hand, plasma from Type II vWD affected patients showed markedly disparent values, with increased (greater than 8.0) vWFAg:CBA ratios coincident with virtually absent CBA binding in these patients. Thus, the CBA as reported here does appear to constitute a novel functional assay capable of detecting qualitative vWF differences in plasma of affected vWD patients.(ABSTRACT TRUNCATED AT 250 WORDS)
Small subsets of central neurons possessing Ca2+ permeable AMPA/kainate channels can be identified by a histochemical stain based on kainate-stimulated Co2+ uptake (Co2+(+)neurons) and are unusually vulnerable to AMPA/kainate receptor-mediated injury. Using brief kainate exposures (which selectively destroy Co2+(+) neurons) along with kainate triggered 45Ca2+ influx measurements, we estimate kainate to cause an unusually high rate of Ca2+ influx into Co2+(+) neurons. Also, while fura-2 Ca2+ imaging revealed low (10 microM) kainate exposures to preferentially induce intracellular free Ca2+ ([Ca2+]i) elevations in Co2+(+) neurons, intense (100 microM) kainate exposures used in the 45Ca2+ influx studies triggered comparable [Ca2+]i rises in all neurons. These findings suggest that the exceptional vulnerability of Co2+(+) neurons to AMPA/kainate receptor-mediated injury reflects a high rate of agonist triggered Ca2+ influx, and that [Ca2+]i rises may only poorly reflect influx rate.
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The susceptibility of phenotypically CCR5-negative resting CD4 T cells for membrane fusion with a CCR5-specific HIV-1 envelope was analysed using a novel sensitive fusion assay. A very low overall density of CCR5 on T cells expressing high levels of CD4 was shown to be sufficient for HIV envelope-mediated membrane fusion. These findings are relevant to the understanding of how HIV-1 R5 strains enter and replicate in resting CD4 T cells in vivo.
Conjunctival flaps are commonly used to cover the cornea in patients who otherwise can not tolerate a scleral shell. An alternative method of protecting the cornea, oral mucous membrane grafting, is described herein. Ten patients had oral mucous membrane grafting to the cornea. Patients ranged from 5 months to 47 years of age. Diagnoses included partial cryptophthalmia, microphthalmia, congenital orbital fibrosis syndrome, juvenile active ossifying fibroma, and trauma. All patients had poor visual function in the affected eye. Patients were either intolerant of scleral shell wear or had other contraindications to the use of a shell over an unprotected cornea. Following mucous membrane grafting (follow-up, 1-3 years), all patients were successfully fitted with prostheses and obtained good cosmetic results.
By cooling the blood just before returning it to the body, marked reduction in the temperature of the heart, brain, liver, and kidney can be produced easily and with great economy in heat transfer, since the great bulk of the animal or patient, consisting of skin, muscle, and bone, is cooled much more slowly. This results in a marked oxygen saving, due to the direct antimetabolic effect of the cooling. Metabolic acidosis can be avoided and so far no irreversible effects of the cooling have been discoverable. Warming is so greatly facilitated by the heat stored in the nonvital areas that it is not necessary to warm the blood returning from the extracorporeal circuit. The use of safe, low, extracorporeal flows offers a number of possible advantages including great simplicity of cannulation and of the pump oxygenator design and operation.
A locked percutaneous device, the LPD-I (previously described as LPD), is effective in overcoming the problems of skin downgrowth and local tissue infection; however, it can only be implanted at a site providing adequate subcutaneous adipose tissue to attach to the subcutaneous connector of the LPD-I. A modified device, the LPD-II, has been developed that has a thin dome and skin connector but does not have a subcutaneous tissue connector. In addition, a newly designed structure called the skin stop collar (SSC) has been developed. It is positioned just beneath the mesh collar described in the LPD-I to further improve the function of the LPD-II. Six rabbits were implanted with one LPD-II without the SSC (group 1) and five rabbits were implanted with one LPD-II with the SSC (group 2). For more than 6 months, two of the implants in group 1 rabbits were successful. Four of the implants in group 2 rabbits were successful for more than 1 year. One of the animals in group 2 died of causes unrelated to the device. We conclude the following: the mesh collar skin connector can function well as a locked percutaneous device without the subcutaneous tissue connector; the LPD-II can be implanted in any site and does not require the presence of subcutaneous adipose tissue; and the SSC may increase the success rate of LPD-II implantation provided the mesh collar is made of soft material.
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The clinical course of a patient presenting with thrombocytopenia (86 X 10(3)/L) and signs of intravascular coagulation (prothrombin time, 45%; partial thromboplastin time, 49 s; fibrinogen, 40 mg/dl; antithrombin III, 85%; factor X, 73%; plasminogen, 42%) due to a giant hemangioma of the liver (Kasabach-Merritt syndrome) is reported. Treatment with i.v. heparin, fibrinogen, and fresh-frozen plasma led to significant elevation of fibrinogen (156 mg/dl) and antithrombin III (102%) without changing the decreased activities of the procoagulant factors. Similarly, the signs of hyperfibrinolysis persisted (fibrinogen degradation products, 50 micrograms/dl; plasminogen, 43%; alpha-2 antiplasmin, 74%). The hemangioma was completely removed by excision of the left lobe of the liver. Subsequently, all coagulation parameters returned to normal, indicating a complete reversibility of the coagulation disorder.
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Sequential immunocytochemical double labeling of glial fibrillary acidic protein (GFAP) and transferrin (Tf) was used for the simultaneous identification of astrocytes and oligodendrocytes in paraffin-embedded sections of the rat brain. Paraformaldehyde or formalin-alcohol-acetic acid fixation followed by protease treatment of paraffin-embedded sections provided the most satisfactory double-labeling. The results demonstrated GFAP-positive astrocytes and Tf-positive oligodendrocytes as two subpopulations of glia with no overlap. Our results suggest that, with some exceptions, immunocytochemical double labeling of GFAP and Tf has a potential application for the simultaneous identification of astrocytes and oligodendrocytes in the rat brain.