Objectives: Plasma exchange or dual-filtration plasmapheresis (DFPP) is used to remove anti-A/anti-B antibodies in ABO-incompatible kidney transplant recipients. However, the substitution fluid used ...for DFPP has a low osmotic pressure, which may cause a decrease in the blood pressure and discomfort due to a decrease in the circulating blood volume during treatment. The aim of the present study was to determine whether DFPP can be performed more safely by adding sodium chloride (NaCl) to the substitution fluid to avoid decrease of the circulating blood volume during treatment. Methods: The study was conducted in 32 patients who needed elimination of anti-A/anti-B antibodies prior to renal transplantation. The first DFPP was performed using 800 ml to 1 L of conventional substitution fluid (a mixture of lactic solution and 20% albumin solution) (control treatment). The second DFPP was performed with substitution fluid to which 20 ml of 10% NaCl had been added (NaCl treatment). Changes in the circulating blood volume (⊿BV) in both treatments were measured every 30 minutes.The blood osmolality and total protein (TP), sodium and hemoglobin (Hb) concentrations were also measured before and after the DFPP. Results: In the NaCl treatment, the osmolality of the substitution fluid increased from 194±12 to 266±18 mOsm. In addition, the ⊿BV was less pronounced from 120 to 210 minutes (P<0.01). The NaCl treatment prevented significant post-treatment plasma osmotic pressure decrease, TP decrease, and Hb increase observed in the control treatment; however, the plasma Na concentration did not change in either treatment. The NaCl treatment was associated with a less pronounced decrease of the blood pressure and less discomfort. Conclusions: Addition of NaCl to the substitution fluid prevented a significant drop in the circulating blood volume during DFPP.
Cold transport of epididymides from genetically modified mice is an efficient alternative to the shipment of live animals between research facilities. Mouse sperm from epididymides cold-stored for ...short periods can maintain viability. We previously reported that cold storage of mouse epididymides in Lifor® perfusion medium prolonged sperm motility and fertilization potential and that the sperm efficiently fertilized oocytes when reduced glutathione was added to the fertilization medium. Cryopreservation usually results in decreased sperm viability; an optimized protocol for cold storage of epididymides plus sperm cryopreservation has yet to be established. Here, we examined the motility and fertilization potential of cryopreserved, thawed (frozen-thawed) sperm from previously cold-stored mouse epididymides. We also examined the protective effect of sphingosine-1-phosphate (S1P) on sperm viability when S1P was added to the preservation medium during cold storage. We assessed viability of frozen-thawed sperm from mouse epididymides that had been cold-transported domestically or internationally and investigated whether embryos fertilized in vitro with these sperm developed normally when implanted in pseudo-pregnant mice. Our results indicate that frozen-thawed sperm from epididymides cold-stored for up to 48h maintained high fertilization potential. Fertilization potential was reduced after cold storage for 72h, but not if S1P was included in the cold storage medium. Live pups were born normally to recipients after in vitro fertilization using frozen-thawed sperm from cold-transported epididymides. In summary, we demonstrate an improved protocol for cold-storage of epididymides that can facilitate transport of genetically engineered-mice and preserve sperm viability after cryopreservation.
Cryopreservation of 2-cell embryos is an effective technology for storage of genetically engineered mouse strains. Transport of genetically engineered mice between laboratories has frequently been ...performed using such cryopreserved 2-cell embryos. However, the receiving laboratory requires proficient skills and special instruments to obtain live young from cryopreserved and transported embryos. Therefore, in this study, we tried to address the storage and transport of vitrified/warmed 2-cell embryos at a cold temperature. In cold storage experiments, the development rates of 2-cell embryos stored in M2 medium for 24, 48 and 72
h into blastocysts were relatively high (83%, 63% and 43%, respectively). Although, 2-cell embryos stored in PB1 and mWM maintained the developmental potency for 24
h, the rates were markedly decreased to low levels after 48
h (PB1: 0%; mWM: 5%). In transport experiments, many pups were obtained from vitrified/warmed 2-cell embryos transported at a cold temperature in all receiving laboratories (incidence of successful development: 49%; 249/511). In summary, short-term storage and transport of vitrified/warmed 2-cell embryos in M2 medium at a cold temperature can maintain their ability to develop into live young.
The C57BL/6 mouse is commonly used to produce transgenic and knockout strains for biomedical research. However, the motility and fertility of its sperm decrease markedly with freezing. Short-term ...preservation of sperm without freezing can avoid this. Furthermore, such samples can be transported safety without the special skills or equipment needed for the transportation of live animals or frozen products. We evaluated the motility and fertility of sperm collected from cauda epididymides after preservation or transportation at 4 super(o)C. Oocytes with the zona pellucida subjected to laser-microdissection were used to assist fertilization in vitro. Although the motility of sperm gradually decreased with storage (P<0.05), no disruption of the sperm plasma membrane was seen. The proportion of zona-intact oocytes fertilized with sperm preserved for 0, 24, 48 and 72h were 70, 14, 5 and 1%, respectively. On the other hand, 45, 20 and 14% of laser-microdissected oocytes were fertilized by sperm preserved for 24, 48 and 72h, respectively (P<0.05). The fertility of sperm collected from cauda epididymides of two transgenic strains after transportation at 4 super(o)C were also significantly increased using laser-microdissected oocytes rather than zona-intact oocytes (57 and 68% vs. 5%, P<0.05). Efficient production of offspring from sperm preserved or transported at 4 super(o)C was achieved using laser-microdissected oocytes. Thus the fertility of sperm preserved or transported at 4 super(o)C could be maintained, although motility gradually decreased with storage. Laser-microdissected oocytes will contribute to the efficient production of embryos and offspring using such preserved sperm samples.
Chronic renal failure patients usually develop secondary hyperparathyroidism and, as the disease progresses, there is a decrease in the number of vitamin D and calcium‐sensing receptors (CaRs) in the ...parathyroid glands. Parathyroid cell function can be controlled if a functional gene is transferred into these cells using an adenovirus vector. Vitamin D or CaR genes transferred by the infected adenovirus vector induced a reduction in parathyroid hormone secretion. These results suggest that adenovirus‐mediated gene transfer is a useful technique for control of parathyroid cell function.
An increased number of blood access troubles such as stenosis, pseudoaneurysms, seroma and thrombosis has been treated. To diagnose the hemodialysis blood access troubles, color Doppler ...ultrasonography was deployed. In 35 patients with chronic renal failure who were sent to our institution for reconstruction of an arteriovenous fistula, color Doppler ultrasonography was performed and the usefulness of the device was evaluated. Stenoses of arteriovenous dialysis grafts were detected most frequently at the venous anastomoses or in native veins proximal to the grafts. Pseudoaneurysms at the puncture sites were presented on the grafts, and exhibited bidirectional flow. No color flow imaging was shown in the seromas. Ultrasound color flow imaging is useful for the exact diagnoses of hemodialysis blood access troubles.
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