A simple system for extending refrigerated, nonfrozen preservation of biological material using pressure.

High pressure (above 238 atmg) substantially extends the refrigerated storage of highly perishable biological material in a nonfrozen state.

5. Calculated Equivalent Values of Pressure and Temperature for Reducing Reaction Rates of Horseradish Peroxidase

Pressure (atmg)	Temperature (°K)	$\text{T}\text{T}{}_0$a
0	296	0.999
272	289	0.975
340	287	0.969
408	285	0.963

a

$\text{T}{}_0\text{= 296 °}\text{K}$It has been known for some time that high pressure stops microbial growth. The effect of high pressure is to reduce further the enzyme activity at refrigerated temperatures. Two enzymes studied, peroxidase and crude trypsin from red crab intestine, demonstrated this effect.A number of food materials such as fish, beef, and chicken were tested for microbial growth and organoleptic qualities after high-pressure storage in a simple 14-liter pressure chamber. Pressure was generated by a hand pump. The results indicated that after 30 days those items held in a non-frozen state at ?3 °C and 238 atmg were not significantly different microbiologically and organoleptically from frozen controls at atmospheric pressure and ?20 °C.This system should be useful for the preservation of biological materials where freezing or thawing effects are undesirable or unknown.The energy saved compared to freezing should also be considered. Only 62% of the energy is required for storage at ?3 °C as compared with frozen storage at ?20 °C, and about 28 cal/g must be removed in cooling to ?3 °C as compared with 120 cal/g in cooling to ?20 °C.

     

Hemopoietic activities of cryopreserved murine fresh and postmortem bone marrow cells

The percentage of preservation of erythropoietic and granulopoietic precursor cells in the murine bone marrow was studied using in vitro methylcellulose clonal cell culture assays and in vivo murine spleen colony assays. This study clearly demonstrates

a. Type of Spleen Colonies Induced by 6-hr Postmortem Murine Bone Marrow Cells^a

	Mean (%)
Type of colonies	t Score	P Value	Unfrozen	Frozen
			Erythrocytic	26.283	14.100	2.09	0.059
Granulocytic	23.741	32.917	1.45	0.173
Mixed	49.321	52.700	0.55	0.59

a

$\text{N = 92}$. the presence of pluripotent hemopoietic precursor cells in cryopreserved 0-, 3-, 6-, 9-, and 12-hr postmortem murine bone marrow cells. Apparently, the erythropoietic precursor cells are more sensitive to freezing injury as compared to granulopoietic precursor cells.

     

Reproductive potential and endocrinological responses of sheep kept under controlled lighting. I. Comparative reproductive performance of four breed types of ewe

Five ewes of each of four breed types were kept in each of two environmentally-controlled rooms over a period of 2.5 years. The daylength varied between 20 and 9 h on a 6-monthly cycle, including a period of 22 weeks during which daylength was decreased by 0.5 h per week, 2 weeks in which it increased from 9 to 20 h, and 2 weeks at 20 h; each room operated 3 months out of phase with the other. Towards the end of the period of decreasing daylength each ewe was synchronised with a progestagen sponge for 12 days, given an injection (i.m.) of 750 I.U. pregnant mares' serum gonadotrophin (PMSG) on withdrawal, and inseminated 48 and 58 h later (500 million fresh undiluted sperm per insemination). Thus, insemination occurred at a single synchronised oestrus every 6 months. Progesterone pregnancy diagnosis was performed on blood samples taken 18 days later and parturition was induced by an injection (i.m.) of 16 mg dexamethasone on day 143 of gestation. Lambs were weaned at birth, allowing the ewes approximately 5.5 weeks to recover before the next insemination and a ‘successive’ conception.The performance of the four types of ewe, starting as maidens, is tabulated.

     

4.

DNA synthesis catalysed by endogenous templates and DNA-dependent DNA polymerases in spermatogenic cells from rat     

Norman B. Hecht  Martti Parvinen 《Experimental cell research》1981,135(1):103-114

The activity levels of DNA polymerases α and β have been measured by autoradiography in squash preparations from rat testis of sexually mature animals. Similar results were obtained with ‘fixed’ samples (dipped in acetone: ethanol for 5 min at 25 °C) or ‘unfixed’ samples (frozen in liquid nitrogen and freeze-dried). The activities of DNA polymerases α and β in situ were distinguished by differential assay conditions and by selective inhibition with compounds such as N-ethylmaleimide and aphidicolin. Using the endogenous chromatin as template, maximal activity for both enzymes was obtained in the presence of all four deoxyribonucleoside triphosphates, MgCl₂ and ethylene glycol. When DNA polymerase activities in several predominant testicular cell types (pre-leptotene primary spermatocytes, pachytene primary spermatocytes, round spermatids and elongated spermatids) were quantitatively compared, on a per cell basis, the following percentage distribution was observed:

Conceptions	Merino	Dorset Horn × Merino	Border Leic. × Merino	South Suffolk × Merino	$\text{P}$
Successive (%)	20.0	12.5	11.1	47.6	$\text{< 0.01}$
Non-successive (%)	76.0	76.9	66.7	95.0	$\text{< 0.05}$
Overall (%)	51.1	52.4	44.4	70.7	$\text{< 0.05}$
Mean litter size	1.7	1.5	1.5	2.2	$\text{< 0.05}$
Mean lambs per ewe per year	1.8	1.6	1.3	3.1	$\text{< 0.01}$

     

5.

Integrity of lysosomes in the isolated perfused rat liver before and after exposure to dimethylsulphoxide.     

D Lee  R K Holland  A K Groufsky 《Cryobiology》1979,16(1):18-23

We have studied the integrity of lysosomes in isolated rat livers perfused for 3, 4, or 6 hr at 35 °C with BSA (40 g/l) in Krebs Ringer bicarbonate buffer. The latency and sedimentability of β-glucuronidase in homogenates of these livers was well maintained even after 6 hr. The latency and sedimentability of acid phosphatase remained at about control levels during the first 4 hr of perfusion but decreased between 4 and 6 hr. These decreases in latency and sedimentability correlated with a decrease in bile production and an increase in the rate of release of GOT into the perfusate and could indicate either intracellular disruption of lysosomes

Latencies, Sedimentabilities, and Specific Activities of Acid Phosphatase and β-Glucuronidase in Homogenates of Rat Liver Prepared before or at Various Times after Exposure to 1.4 m Me₂SO for 1 hr

	Pre-leptotene primary spermatocyte %	Pachytene primary spermatocyte %	Round spermatid %	Elongated spermatid %
DNA polymerase α	25	42	30	3
DNA polymerase β	29	34	36	1

     

6.

An experimental study using tissue culture medium as preservation and perfusion fluid     

T Nilsson  E Schueller  G P Murphy  W J Staubitz 《Cryobiology》1973,10(3):191-196

A commercially available tissue culture medium has been proven capable of preserving dog kidney function for at least 24 hr after simple cooling. The advantages of using tissue culture medium as preservation fluid instead of plasma or albumin solutions from the infectious and immunological points of view are obvious. An in vitro study was completed using the tissue

1.

Time (hr)	0	3	4	6
Acid phosphatase
Latency (%)	83.2 ± 0.8	64.7 ± 5.1	62.4 ± 7.9	68.9 ± 5.4
Sedimentability (%)	81.7 ± 0.6	77.6 ± 3.1	81.0 ± 3.4	79.4 ± 5.1
Specific activity (mIU/mg protein)	2.8 ± 0.4	2.8 ± 0.2	2.4 ± 0.4	1.6 ± 0.2
β-Glucuronidase
Latency (%)	06.8 ± 1.5	71.3 ± 4.3	74.2 ± 2.5	63.3 ± 4.5
Sedimentability (%)	69.5 ± 0.3	75.4 ± 3.2	75.0 ± 1.1	74.6 ± 2.0
Specific activity (mIU/mg protein)	1.8 ± 0.1	1.6 ± 0.1	1.6 ± 0.2	1.6 ± 0.2

     

7.

A digital thermocouple thermometer     

A R Hayes 《Cryobiology》1974,11(4):378-381

Measurements of the reproducibility of a random selection of copper/constantan thermocouples were made and it was found that they agreed within 1 ° C. Based on this finding, a digital thermocouple thermometer was designed and constructed incorporating a thermocouple linearizer and cold junction compensation. The instrument

Accuracy of the Completed Digital Thermometer

Code (animal No.)	Perfusion time (br)	Perfusion pressure mm/Hg	Flow ml/min	Weight gain	pH	pO2 mm/Hg	Histological appearance
1	24	70-60 systolic	96	35	7.3	150–180	Grossly normal
2	24		45-40 diastolic	108	30
3	24		96	30
4	48	70-60 systolic	80	35	7.3	150–180	Grossly normal
5	48	45-40 diastolic	120	40	7.4
6	48		100	40
7	72	70-60 systolic	115	40	7.4	150–180	Slight vacuolization of the tubular cells
8	72	45-40 diastolic	96	40
9	72		80	40
10	24	70-60 systolic	110	35	7.3	150–180	Used for transplantation
11	24	45-40 diastolic	120	35
12	24		140	40
13	24		100	30
14	24		96	30

     

8.

Effects of changes in sodium electrochemical potential gradient on p-aminohippurate transport in newt kidney     

Yoshikatsu Kikuta  Hiromu Hayashi  Yoshitaka Saito 《生物化学与生物物理学报:生物膜》1979,556(2):354-365

     

9.

Purification and characterization of ($\text{Na}{}^{\mathrm{+}}\text{+ K}{}^{\mathrm{+}}$)-ATPase. VI. Differential tryptic modification of catalytic functions of the purified enzyme in presence of NaCl and KCl     

Peter Leth Jørgensen 《生物化学与生物物理学报:生物膜》1977,466(1):97-108

     

10.

Cell Biology: Paper alert     

《Current opinion in cell biology》2000,12(5):525-535

A selection of interesting papers that were published in the two months before our press date in major journals most likely to report significant results in cell biology.

Temperature	Indicated	Error
(°C)	temperature	(°C)
	(°C)
?1.95.75	?195	0.75
?77.02	?78	0.38
0	0	0
52.49	53	0.51
Mean		0.413

     

11.

Monovalent cation activated ouabain-insensitive ATPase in the gills of freshwater mussel Anodonta cygnea     

《Comparative biochemistry and physiology. B, Comparative biochemistry》1992,101(4):903-908

     

12.

Heat exchange in the cryosurgery of Menière's disease: Experimental and clinical studies     

S. Harly  J.E. Aastrup  O. Elbrønd 《Cryobiology》1977,14(5):609-613

The temperature course in the lateral semicircular canal and in the facial canal was studied in experiments during freezing of the semicircular canal. The course of the temperature was measured with thermocouples. Concurrently, the heat flow was measured, and also the total heat exchange was measured throughout the freezing period by a thermoelectric heat flowmeter incorporated in the cryotip. The measurements showed correlation between the total amount of heat exchanged, the freezing time, and the temperature in the semicircular canal. This correlation was utilized to assess and calculate (the temperature of the lateral semicircular canal) the course of the cryoprocess in vivo, where it is possible to measure the heat flow and the total heat exchange during the freezing period only.

2. Results upon Vertigo

Contents (chosen by)
525	Cytoskeleton (Desai and Holleran)
526	Cell regulation (Roche, Servant and Weiner)
528	Nucleus and gene expression (Aasland and Weinzierl)
529	Membranes and sorting (Ponnambalam)
530	Membrane permeability (Slesinger)
531	Cell-to-cell contact and extracellular matrix (Pfaff)
533	Cell differentiation (van Roessel, Kaltschmidt, Tsang and Huckriede)
534	Cell multiplication (Sclafani)

     

13.

Studies on $\text{(Na}{}^{\mathrm{+}}\text{+ K}{}^{\mathrm{+}}\text{)-activated}$ ATPase. XLVI. Effect of cation-induced conformational changes on sulfhydryl group modification     

B.M. Schoot  S.E. Van Emst-De Vries  P.M.M. van Haard  J.J.H.H.M. de Pont  S.L. Bonting 《生物化学与生物物理学报:生物膜》1980,602(1):144-154

     

14.

Influence of adrenalectomy upon rat erythrocyte Na⁺ and K⁺ content,Na⁺ efflux rate and Mg²⁺- and ($\text{Na}{}^{\mathrm{+}}\text{+}\text{K}{}^{\mathrm{+}}$)-Mg²⁺-ATPase activities     

Michael A. Radcliffe 《生物化学与生物物理学报:生物膜》1974,339(3):303-310

     

15.

Lysophosphatidic acid as a stimulator of Na+-dependent Ca2+ efflux from adrenal chromaffin cells     

《Life sciences》1995,57(15):PL205-PL210

     

16.

Isolation of the basal and lateral plasma membranes of rat kidney tubule cells     

H. Ebel  E. Aulbert  H.J. Merker 《生物化学与生物物理学报:生物膜》1976,433(3):531-546

     

17.

A theoretical analysis of the effect of phosphate on apparent H+O stoichiometries in oxygen-pulse experiments with rat liver mitochondria     

《BBA》1986,849(2):236-243

     

18.

In vivo effects of copper on the calcium homeostasis mechanisms of mussel gill cell plasma membranes     

《Comp. Biochem. Physiol. C, Comp. Pharmacol. Toxicol.》1996,113(3):421-425

     

19.

Study of the differential modifications of ($\text{Na}{}^{\mathrm{+}}\text{+ K}{}^{\mathrm{+}}$)-ATPase and its partial reactions by dimethylsulfoxide     

Kenichi Kaniike  Erland Erdmann  Wilhelm Schoner 《生物化学与生物物理学报:生物膜》1974,352(2):275-286

     

20.

Adrenomedullin stimulates calcium efflux from adrenal chromaffin cells in culture: Possible involvement of an Na+Ca2+ exchange mechanism     

《Life sciences》1995,58(2):PL35-PL40

     

	No Vertigo	Improved	Unchanged
Number of patients	7	5	3

DNA synthesis catalysed by endogenous templates and DNA-dependent DNA polymerases in spermatogenic cells from rat

The activity levels of DNA polymerases α and β have been measured by autoradiography in squash preparations from rat testis of sexually mature animals. Similar results were obtained with ‘fixed’ samples (dipped in acetone: ethanol for 5 min at 25 °C) or ‘unfixed’ samples (frozen in liquid nitrogen and freeze-dried). The activities of DNA polymerases α and β in situ were distinguished by differential assay conditions and by selective inhibition with compounds such as N-ethylmaleimide and aphidicolin. Using the endogenous chromatin as template, maximal activity for both enzymes was obtained in the presence of all four deoxyribonucleoside triphosphates, MgCl₂ and ethylene glycol. When DNA polymerase activities in several predominant testicular cell types (pre-leptotene primary spermatocytes, pachytene primary spermatocytes, round spermatids and elongated spermatids) were quantitatively compared, on a per cell basis, the following percentage distribution was observed:

Integrity of lysosomes in the isolated perfused rat liver before and after exposure to dimethylsulphoxide.

We have studied the integrity of lysosomes in isolated rat livers perfused for 3, 4, or 6 hr at 35 °C with BSA (40 g/l) in Krebs Ringer bicarbonate buffer. The latency and sedimentability of β-glucuronidase in homogenates of these livers was well maintained even after 6 hr. The latency and sedimentability of acid phosphatase remained at about control levels during the first 4 hr of perfusion but decreased between 4 and 6 hr. These decreases in latency and sedimentability correlated with a decrease in bile production and an increase in the rate of release of GOT into the perfusate and could indicate either intracellular disruption of lysosomes

Latencies, Sedimentabilities, and Specific Activities of Acid Phosphatase and β-Glucuronidase in Homogenates of Rat Liver Prepared before or at Various Times after Exposure to 1.4 m Me₂SO for 1 hr

An experimental study using tissue culture medium as preservation and perfusion fluid

A commercially available tissue culture medium has been proven capable of preserving dog kidney function for at least 24 hr after simple cooling. The advantages of using tissue culture medium as preservation fluid instead of plasma or albumin solutions from the infectious and immunological points of view are obvious. An in vitro study was completed using the tissue

1.

A digital thermocouple thermometer

Measurements of the reproducibility of a random selection of copper/constantan thermocouples were made and it was found that they agreed within 1 ° C. Based on this finding, a digital thermocouple thermometer was designed and constructed incorporating a thermocouple linearizer and cold junction compensation. The instrument

Accuracy of the Completed Digital Thermometer

Cell Biology: Paper alert

A selection of interesting papers that were published in the two months before our press date in major journals most likely to report significant results in cell biology.

Heat exchange in the cryosurgery of Menière's disease: Experimental and clinical studies

The temperature course in the lateral semicircular canal and in the facial canal was studied in experiments during freezing of the semicircular canal. The course of the temperature was measured with thermocouples. Concurrently, the heat flow was measured, and also the total heat exchange was measured throughout the freezing period by a thermoelectric heat flowmeter incorporated in the cryotip. The measurements showed correlation between the total amount of heat exchanged, the freezing time, and the temperature in the semicircular canal. This correlation was utilized to assess and calculate (the temperature of the lateral semicircular canal) the course of the cryoprocess in vivo, where it is possible to measure the heat flow and the total heat exchange during the freezing period only.

2. Results upon Vertigo