Use of Inert Gases for the Preservation of Nuclear Blood Cells

Abstract The subject of the study was the stability of human white blood cell membranes subject to noble gases (xenon ad krypton, 0.6 mPa) clathrate cryoanabiosis (‒80°C). A unique portable stainless steel low pressure container with a compartment for flexible plastic container was designed to ensure that the cells are saturated with gases. The samples were warmed after 1 and 30 days in a water bath (+38°C) for 35-50 sec, while the container was being tilted (2-3 times per second), until the temperature of the biological object reached +3±1°C. It was demonstrated that after 30 days of clathrate anabiosis (-80°C) over 95% (of the original number) of leukocytes remain viable, and cell membranes of 54.5±3.4% of them is resistant to trypan blue; granulocyte survival rate is 73.5±2.7%, original lipid peroxidation rate and antioxidant activity are retained. Biological object cryopreservation in noble gases environment is a promising trend in biology and medicine.

Alterations of the Ventilation-Perfusion Ratio Measured by Using a Multiple Inert Gas Elimination Technique (MIGET) in One Lung Ventilated Rabbit / 대한마취과학회지

BACKGROUND: In this study, we performed one-lung ventilation (OLV) in rabbits to assess the effects of OLV on the VA/Q ratio and the respiratory physiological changes using MIGET. METHODS: Ten male New Zealand white rabbits, weighing 3-4 kg were selected. To perform MIGET, six inactive gases (SF6, krypton, desflurane, enflurane, diethyl ether, acetone) in 500 ml normal saline were injected intravenously. During two-lung ventilation (TLV), and after OLV for 30 minutes, blood was sampled for blood gas analysis and MIGET, hemodynamic variables were measured. For MIGET, the concentrations of the injected inert gases were measured and converted to retention/excretion data; the VA/Q distribution curve was obtained using a computer. RESULTS: Systolic, mean, and diastolic pulmonary pressures were elevated significantly and pulmonary resistance was doubled (P<0.05) in OLV compared to TLV. Blood pH decreased in OLV. The calculated intrapulmonary shunt was 19% and 52%, TLV and OLV, respectively. The analysis of VA/Q using MIGET showed that the VA/Q distribution curve was wider and that the VA/Q area was larger in normal rabbits. And, that intrapulmonary shunt approximated to 11%. In the case of OLV, a significant increase in shunt was observed but no change in the amount of dead space at distribution area, (log SDQ, log SDV) remained the same, whereas the VA/Q distribution curve shifted toward the right. CONCLUSIONS: OLV in rabbits showed severe hypercapnia and hypoxemia leading to a considerable increase in shunt. Because of the wide range of VA/Q distribution in TLV, no significant changes in respiratory variables were observed during OLV.

Measurement of Ventilation-Perfusion Ratio Using Multiple Inert Gas Elimination Technique in the Endotoxin-Induced Septic Rabbit / 대한마취과학회지

BACKGROUND: Endotoxin is a complex lipopolysaccharide molecule situated within the outer membrane of Gram-negative bacteria. Sepsis and acute respiratory failure (ARDS) can be induced by endotoxin. In order to introduce and develop the experimental model of ARDS in sepsis, we induced sepsis with the endotoxin and investigated the change of respiratory pathophysiology during sepsis using a multiple inert gas elimination technique (MIGET). METHODS: Ten New Zealand white rabbits were anesthetized and ventilated with a Harvard apparatus. In 5 rabbits, 2 mg/kg of lipopolysaccaride from E. coli was infused intravenously for 30 min (Toxin group). At 1, 2, 3, and 4 hours after endotoxin infusion, arterial blood gas, and hemodynamic profiles were checked. To perform the MIGET, six inert gases (SF6, krypton, desflurane, enflurane, diethyl ether, acetone) of widely varying solubility were infused peripherally and the excretion and retention data was determined from measurements of inert gas tensions in pulmonary arterial, systemic arterial blood samples and mixd expiratory gas sampling of pre and post septicemia using gas chromatography. We transformed and analysed the data into a V/Q distribution curve to find out the change of V/Q distribution curve. After the experiments, the animals were dissected and the lungs were extracted for wet/dry weight ratio (WW/DW) and microscopic examination. RESULTS: In the Toxin group, the pulmonary arterial pressures were increased and arterial oxygen tensions were decreased after the endotoxin infusion. The lung WW/DW were increased and inflammatory findings were seen in microscopic examination. In the MIGET, shunt, deadspace and log SDQ were increased in the toxin group, though there were wide V/Q distributions in the control group. CONCLUSIONS: We developed a successful endotoxin induced septic animal model, V/Q distribution curve and data using MIGET. The accomplishment of the experiment will not only allow us to better understand pulmonary pathophysiology of endotoxin induced sepsis using MIGET, but it will also contribute to other pulmonary physiology experiments associated with sepsis.

Neurotropic Effect of Basic Fibroblast Growth Factor on Laser Wound

In order to analyze the potential role of basic fibroblast growth factor (bFGF)on retinal wound after laser photocoagulation, we studied the expression of GFAP and the morphologic changes in the retina of pigmented rats. A multiline krypton laser was used to produce laser lesions. After irradiation, eyes were intravitreally injected with 1 microliter vehicle or 1 microgram of bFGF in l microliter of vehicle. And we examined the expression of GFAP at 1 day, 3 days, 7 days and 14 days after treatment.We also measured the size of defect in outer nuclear layer at 7 days after laser. The results were as follows : 1. GFAP immunoreactivity of M ü ller cell begins to appear within 24 hours in whole retina area.It sharply increases until 7 days and then decreases slowly. 2. bFGF stimulates the expression of GFAP and it is statistically significant(p<0.05) 3. BFGF prevents cell death of photoreceptor in the laser lesions(p<0.05 ) From these results, we concluded that bFGF stimulates Müller cell reactivity after laser phtocoagulation and prevents photoreceptor loss. These results may be important to understand the healing mechanism of laser phtocoagulation and to minimize the complications of laser phtocoagulation.

A Comparison of Cyclocryotherapy and Contact Transscleeral Cyclophotocoagulation with Diode Laser in Rabbit

Cyclodestruction is generally achieved by applying thermal energy across the intact sclera. Commonly employed modalities are ultrasound, cryopexy, diathermy, noncontact laser light and most recently, contact transscleral laser using argon, krypton and Nd:YAG, diode laser. This study was performed to compare the pressure lowering and tissue effects of contact transscleral cyclophotocoagulation(TSCPC) with diode laser using G prode to cyclocryotherapy in rabbit. The results of this study demonstrate a similar intraocular pressure lowering effect of both cyclocryotherapy and contact TSCPC with diode laser using G prode. Histologic studies showed that both TSCPC and cyclocryotherapy are effective anatomic cyclodetructive modalities but in case of TSCPC, less severe destruction of ciliary body was noted. This study indicated that contact TSCPC with diode laser using G probe delivery provieds effective and safe management for eyes with medically and surgically uncontrolled glaucoma compared to cyclocryotheraphy.

Peripheral Scatter Treatment in Diabetic Retinopathy

Panretinal photocoagulation (PRP) has been known to be beneficial in preventing severe visual loss from diabetic retinopathy. Clinical effects of peripheral scatter treatment on diabetic retinopathies were compared with those of PRP in 40 eyes of each groups. In peripheral scatter treatment, equatorial and peripheral fundus was treated with argon blue-green laser or krypton red laser, in 2 sessions with interval of one week. The incidence of maculopathy causing decrease in visual acuity after laser treatment was higher in PRP-treated group than in peripheral scatter treatment group, but changes in visual acuity and fundus findings at last follow-up were not significantly different between PRP-treated and peripheral scatter treatment group (p>O.05). Peripheral scatter treatment was equally affective as PRP in treating diabetic retinopathy with less amount of laser bums and saving post-equatorial fundus.

Levaduras aisladas de mostos de uva en la zona de Oltrepo Pavese, Italia, y su sensibilidad in vitro frente a compuestos de cobre y zinc / Yeast isolated from musts of grapes in Oltrepó Pavese, Italy, and their in vitro sensibility to zinc and copper compounds

Se investigó durante el periodo 1989-90 en la región de Oltrepó Pavese (Italia), la micota de los mostos provenientes de uvas negras y blancas. De un total de 28 especies levaduriformes aisladas, incluidas en 10 géneros, Saccharomyces, fué el género dominante con las especies S.italicus y S.cerevisiae. Tambien se detectaron frecuentemente levaduras anascosporógenas, tales como Candida valida, Torulopsis (C) holmii y Kloeckera apiculata. Se efectuaron estudios de sensibiliadad in vitro frente a compuestos de CuSO4, ya sea en cepas de S. cerevisiae aisladas de los mostos, como de Alternaria, Botrytis, Cladosporium y Sporobolomyces, aisladas del filoplano de la uva. Las concentraciones de CuSO4 a 0.007M y a 0,062M, fueron fúngicidas para S.cerevisiae y Sporobolomyces roseus respectivamente

Clinical Evaluation of Subretinal Neovascular Membrane

Subretinal neovascular membrane in the macula is a major cause of central visual loss. We reviewed the clinical records, fundus photography, fluorescein angiography of the 172 eyes of 166 patients in order to disclose the causes and clinical characteristics of subretinal neovascular membrane and performed argon and krypton photocoagulation in 22 patients. The common causes of subretinal neovascular membrane are idiopathic neovascular membrane, myopic and senile macular degeneration in order(27.1, 26.5, 22.3% aa). There was no significant difference in changes of visual acuity and extent of subretinal neovascular membrane between treatment and non-treatment groups.

A Case of Bilateral Central Serous Chorioretinopathy in a Chronic Tibial Osteomyelitis Patient

Central serous chorioretinopathy (CSC) is characterized by central serous retinal detachment. It occurs usually in young adult male, and bilateral involvement is uncommon. We have experienced bilateral CSC occurred in a 33 year old male patient with chronic tibial osteomyelitis. We treated only the right eye with krypton laser to compare the course of laser treated eye with untreated opposite eye. There was no difference between the course of two eyes in our case.

A Case of Treatment of Macular Subretinal Neovascularization with the Red-Light Krypton Laser

A case of macular subretinal neovascular membrane was treated by red-light krypton photocoagulation. We used continuous red-light krypton laser(647.1nm) because this light is not absorbed by the hemoglobin in the retinal hemorrhage or by the macular luteal pigment(Xanthophyll). 3 weeks after treatment there was no specific change in the treated area. 6 weeks later, we found that the treated area showed some regression of subretinal neovascular membrane. After 3 monthes, the destruction of the subretinal neovascular membrane was complete, and the visual acuity was improved.