Ginkgolide B production in cultured cells derived from Ginkgo biloba L. leaves

Callus cultures and cell suspension cultures derived from Ginkgo biloba L. leaves produced ginkgolidc B. In cell suspension cultures, the production reached a maximum by the 13th day of subculture and followed by a sharp decrease. The medium of Murashige and Skoog induced the highest ginkgolide B content in cultures while the medium of Schenk and Hildebrandt promoted cell growth. For the maximal production of ginkgolide B, cells were cultured in Murashige and Skoog medium modified to contain 1.0 mg/l of -naphthaleneacetic acid, 0.1 mg/1 of kinetin, 30 g/1 sucrose and 1.25 mM potassium phosphate with a molar ratio of ammonium to nitrate ions of 1 3.Abbreviations B5 Gamborg et al (1968) medium - GKB Ginkgolide B - MS Murashige and Skoog (1962) medium - NAA -naphthaleneacetic aicd - SH Schenk and Hildebrandt (1972) medium     

NMR relaxation characteristics of rubidium-87 in perfused rat salivary glands.

Rubidium is a good substitute for potassium in many biological systems, and it has been suggested that rubidium-87 nuclear magnetic resonance (87Rb-NMR) spectroscopy could be used to measure K+ fluxes across membranes in intact tissues. To evaluate this possibility, isolated rat mandibular salivary glands were perfused with solutions containing Rb+ in place of K+. The 87Rb signals arising from the intra- and extracellular compartments were first separated by spectral subtraction and then subjected to line-shape analysis. The narrow extracellular signal was a single Lorentzian (line-width 156 Hz), whereas the broader intracellular signal consisted of two Lorentzian components (ca. 530 and 3080 Hz). Double-quantum filtering of the 87Rb signal from the glands revealed two components of transverse relaxation in antiphase (rate constants 1.8 and 13.3 ms-1), showing the probable involvement of quadrupolar interactions in the relaxation of intracellular Rb+. We conclude, therefore, that both line-shape analysis and double-quantum filtering could provide a basis for the measurement of unidirectional K+ fluxes in intact tissues.     

Chromatographic Evidence for the Presence of Multiple Tachykinins in the Bovine Adrenal Medulla

Among the mammalian tachykinins, substance P (SP) has been shown to be the most potent at modulating the response due to nicotinic acetylcholine receptor stimulation of bovine adrenal chromaffin cells. SP-like immunoreactivity has been detected in nerve terminals innervating the adrenal medulla; however, little is known of the presence of other tachykinins in this tissue. In this study, reverse-phase HPLC was used to fractionate peptides in bovine adrenal medullary extracts, and the fractions were analyzed by radioimmunoassay using antisera to SP or neurokinin A (NKA). The results show that both NKA- and SP-like immunoreactivities are present in the adrenal medulla. The presence of neurokinin B is also indicated. The presence of multiple tachykinins in this tissue raises questions as to their functions in the adrenal medulla.     

Source of prolactin in human follicular fluid.

To analyze whether prolactin (PRL) in human follicular fluid (FF) is synthesized locally or derived from the circulation, PRL concentrations of plasma and FF were determined in the patients after ovarian stimulations. The amounts of PRL messenger ribonucleic acid (mRNA) in the follicular tissues during different menstrual phases were also determined. The FF PRL concentration was correlated positively with plasma PRL and highest estradiol levels during the stimulatory cycle. No PRL mRNA sequence was detected in the RNAs extracted from follicles at any stage in the menstrual cycle, although beta-actin mRNA was detected in all samples. In a comparison with pituitary RNA, the PRL mRNA concentration in ovarian follicular tissues seemed to be 10,000 times less than that in the pituitary. These results suggest that FF PRL may not be synthesized locally, but derived from the pituitary via the circulation through passive diffusion, and thus regulated by estrogen.     

NMR characteristics of intracellular K in the rat salivary gland: a 39K NMR study using double-quantum filtering

Intracellular K of the perfused rat mandibular salivary gland was measured by 39K NMR spectroscopy at 8.45 T. Multiple-quantum NMR arising from multiple-exponential decay was used to eliminate the resonance due to extracellular K in the perfused gland at 25 degrees C. The resonance due to intracellular K consisted of two Lorentzian signals stemming from the [spin 1/2 to -1/2] coherence (sharp resonance) and the [spin -1/2 to -3/2], [spin 3/2 to 1/2] coherences (broad resonance). The transverse relaxation time (T2) corresponding to the [spin 1/2 to -1/2] coherence was ca. 2.5 ms, and that corresponding to the [spin -1/2 to -3/2], [spin 3/2 to 1/2] coherences was ca. 0.4 ms. The relaxation time of the double-quantum coherence of rank 3 (originating from product operators like Ix2Iz) was determined to be ca. 0.2 ms. These results suggest the possibility of the presence of a single homogeneous population of intracellular K with a correlation time of ca. 2.5 x 10(-8) s and a quadrupolar coupling constant of ca. 1.4 MHz.     

Measurement of intracellular Na in the rat salivary gland: a 23Na-NMR study using double quantum filtering

23Na in the prefused rat mandibular salivary gland was measured by spin-echo double quantum filter 23Na-NMR spectroscopy at 8.45 T. Resonances due to the intracellular 23Na and the interstitial 23Na were observed in the perfused gland at 25 degrees C. The resonance due to intracellular 23Na consisted of two Lorentzian signals stemming from the [1/2 mean value of -1/2[ coherence (sharp resonance) and the [-1/2 mean value of -3/2[ and [3/2 mean value of 1/2[ coherences (broad resonance). The transverse relaxation rate constant corresponding to the [1/2 mean value of -1/2[ coherence was 95 +/- 4 s-1 and that corresponding to the [-1/2 mean value of -3/2[ and [3/2 mean value of 1/2[ coherences was 1360 +/- 75 s-1 (mean +/- S.E., n = 5). The resonance due to the interstitial 32Na had longer relaxation rate constants, and disappeared upon administration of dysprosium triethylenetetramine-N,N',N",N",N"'-hexaacetic acid.     

Analysis of E. coli phoA-lacZ fusion gene expression inserted into a multicopy plasmid and host cell's chromosome

The production of beta-galactosidase from the E. coli phoA-lacZ fusion gene was studied to compare the gene expression behavior of two cloning methods: insertion to multicopy plasmids and integration into host cell's chromosome. The chromosome-integrating strain showed more tight control of fusion gene expression levels than the plasmid-containing strain. A 100-fold enhancement of specific beta-galactosidase activity in the former strain was achieved in response to changes of initial inorganic phosphate concentration from 1 to 0.1 mM, whereas a 26-fold increase was observed in the latter strain. The low degree of overexpression in the plasmid-bearing cells was due to a combination of factors including leaky expression in repressed conditions and limitation of biosynthetic machinery in derepressed conditions. In a mixture of inorganic and organic phosphates, inorganic phosphate levels in the medium exhibited oscillatory behavior. The oscillation of inorganic phosphate is attributed to selective usage of inorganic phosphate followed by hydrolysis of organic phosphate to inorganic by alkaline phosphatase. The fluctuation of inorganic phosphate levels also caused the oscillation of beta-galactosidase activity.     

Studies on hemoglobin variants in Korean cattle

814 hemoglobin samples from Korean cattle were investigated by starch gel electrophoresis for the detection of hemoglobin variants. A new variant of cattle hemoglobin, called Hb, H, was recognized. It has a slower rate of migration than Hb A.
Hemoglobin types from 814 Korean cattle were as follows: 652 of Hb AA type (80.1 %), 133 of Hb AB (16.4 %), 12 of Hb AC (1.5 %), 9 of Hb BB (1.1 %), 2 of Hb BC (0.2 %), 4 of Hb AH (0.5 %), 1 of Hb CH (0.1 %), 1 of Hb HH (0.1 %). These figures correspond to the frequencies: Hb^A = 0.893, Hb^B = 0.093, Hb^c =0.009, Hb^H = 0.004.
Gene frequencies at the Hb locus in Korean cattle were compared among six population from different provinces. Generally, high frequencies of Hb^A were observed in each province.     

Spatiotemporal Clusters and Trends of Pneumocystis Pneumonia in Korea

This study determined the recent status and trend of Pneumocystis jirovecii pneumonia (PcP) in the non-human immunodeficiency virus (HIV) (non-HIV-PcP) and HIV (HIV-PcP) infected populations using data from the Health Insurance Review & Assessment Service (HIRA) and the Korea Disease Control and Prevention Agency (KDCA). SaTScan and Joinpoint were used for statistical analyses. Non-HIV-PcP cases showed an upward trend during the study period from 2010 to 2021, with the largest number in 2021 (551 cases). The upward trend was similar until 2020 after adjusting for the population. Seoul had the highest number of cases (1,597) in the non-HIV-PcP group, which was the same after adjusting for the population (162 cases/1,000,000). It was followed by Jeju-do (89 cases/1,000,000). The most likely cluster (MLC) for the non-HIV-PCP group was Seoul (Relative Risk (RR)=4.59, Log Likelihood Ratio (LLR)=825.531), followed by Jeju-do (RR=1.59, LLR=5.431). An upward trend was observed among the non-HIV-PcP group in the Jeju-do/Jeollanam-do/Jeollabuk-do/Gyeongsangnam-do/Busan/Daejeon/Daegu/Ulsan joint cluster (29.02%, LLR=11.638, P<0.001) located in the southern part of Korea. Both women and men in the non-HIV groups showed an overall upward trend of PcP during the study period. Men in the 60–69 age group had the highest annual percentage change (APC 41.8) during 2014–2019. In contrast, the HIV groups showed a falling trend of PcP recently. Men in the 60–69 age group had the most decrease (APC −17.6) during 2018–2021. This study provides an analytic basis for health measures and a nationwide epidemiological surveillance system for the management of PcP.     

Black soldier fly (Hermetia illucens) larvae oil as an alternative fat ingredient to soybean oil in laying hen diets

ObjectiveThe objective of this study was to determine whether dietary black soldier fly (Hermetia illucens, HI) larvae oil (HILO) could serve as an alternative fat source to soybean oil (SBO) in laying hen diets.MethodsWe randomly assigned 25-week-old Hy-line Brown laying hens (n = 144) to receive (n = 6 hens/group; eight replicates) a control or an experimental diet in which SBO was replaced with 50% (50HILO) or 100% HILO (100HILO).ResultsDietary HILO did not negatively affect body weight or productive performance during the study. The eggs also had similar quality parameters, proximate composition, and cholesterol levels. However, the yolk color index was significantly higher (p<0.01) in the 100HILO than in the other groups. Dietary HILO significantly altered the composition of fatty acids (FAs) in abdominal fat and eggs. Total saturated fatty acids (SFAs) and total polyunsaturated FAs (PUFAs) were significantly increased and decreased in the 50HILO and 100HILO groups, respectively, compared with those in the control group (p<0.001 and p<0.0001, respectively). Specifically, the medium-chain FAs lauric and myristic acids were remarkably increased in the abdominal fat of laying hens fed HILO (p<0.0001), whereas only myristic acid increased in eggs (p<0.0001). Undesirable heavy metal (aluminum, fluorine, arsenic, lead, mercury, and cadmium) concentrations were below permissible limits in eggs.ConclusionWe considered that HILO could be an alternative dietary fat to SBO for laying hens with maintained productive performance and good egg quality.