Jar Culture Research Articles

Recent research on the stone jars of Northeast India: Evidence from East Jaintia Hills of Meghalaya

The stone jar sites of Northeast India have been a subject of great interest since their first discovery in 1929 by James Philip Mills and John Henry Hutton in North Cachar Hills, Assam. In Southeast Asia, they were first reported by McCarthy (1900) in Laos and subsequently by the French archaeologist Madeleine Colani in 1935, who reported them in great numbers and speculated about their connections to Assam within a wider salt trade network. The present paper focuses on the stone jar sites from an unreported region of Northeast India along the Saipung Sub-division in East Jaintia Hills in the state of Meghalaya. The stone jars from the sites form the extreme westerly extension of the stone jar culture reported from Northeast India. The results of preliminary fieldwork undertaken in February 2020 across the Saipung reserved forest of Meghalaya led to the discovery and documentation of eight previously unreported sites. A small-scale excavation of four jars in the East Jaintia Hills have helped to provide key insights on the mortuary practices of the people who made and used these stone jars. Material evidences from the excavation clearly suggest that the stone jars are visible relics which were erected on top of a pit where the post-cremated cultural materials of the dead are buried. Such significant findings bear relevance to the further understanding of stone jar sites within the broader context of Northeast India and Southeast Asia.

The comparison of pegylated liposomal doxorubicin and beta-carotene effects on JAR and JEG-3 choriocarcinoma human cell culture models

Objective:The aim was to investigate the effectiveness of pegylated liposomal doxorubicin (PLD), beta-carotene, and a combination of PLD and beta-carotene on JAR and JEG-3 human choriocarcinoma (CC) cell lines for the treatment of CC.Material and Methods:JAR and JEG-3 cells were cultured. PLD and beta-carotene trial groups were determined with different doses (for single drug trial; PLD 1, 2, 5 μg/mL and beta-carotene 1, 5, 10 μg/mL, and for combined drug trial; all PLD doses combined with beta-carotene 5 μg/mL). Drugs were administered to cultures simultaneously, and 72 hours later the cells were detached using trypsin-ethylenediamine tetraacetic acid solution. The percentage of apoptotic cells was determined by flow cytometry after annexin V staining. One set of the supernatant was collected before trypsin application to investigate beta-human chorionic gonadotropin (β-hCG) and hyperglycosylated hCG (H-hCG) levels. Statistical analyses of the apoptotic ratios were performed using Shapiro-Wilk, Kruskal-Wallis and Mann-Whitney U tests.Results:Apoptosis increased in JAR and JEG-3 cultures after treatment with all doses of PLD (p<0.05). A single application of each beta-carotene dose increased apoptosis in JAR cells (p<0.05) but had no apoptotic effects on JEG-3 cells. In the PLD and beta-carotene combination group, apoptosis increased in both JAR and JEG-3 cells (p<0.05).Conclusion:To our knowledge, this is the first investigation of the effectiveness of PLD, beta-carotene, and PLD + beta-carotene combination therapy in two different CC cell lines. PLD is a promising chemotherapeutic drug, and beta-carotene can be used as a novel non-chemotherapeutic agent for treatment of CC. Based on the results of this study, vitamin A supplementation may have promise as a preventive measure. However, these data need support from animal experiments and clinical trials.

Consuming Desire: Sylvia Plath’s The Bell Jar and 1950s American Consumer Culture

Consuming Desire: Sylvia Plath’s The Bell Jar and 1950s American Consumer Culture

Submerged Culture of Grifola gargal and G. sordulenta (Higher Basidiomycetes) from Argentina as a Source of Mycelia with Antioxidant Activity.

Submerged culture is an alternative mycelium source for Grifola gargal and G. sordulenta, two rare edible mushrooms related to Grifola frondosa. This work studies their mycelia as a source of antioxidants. The efficient concentrations of methanolic extracts in both radical scavenging (RS) and reducing power (RP) abilities in G. gargal and in G. sordulenta showed a high antioxidant activity. In the experimental design used, the antioxidant activity mainly depended on the culture conditions rather than on the media composition. Irrespective of the basal culture medium, mycelium methanolic extracts of G. sordulenta obtained from culture in Erlenmeyer flasks showed equivalents to ascorbic acid (EQ(AA)) RS-EQ(AA) and RP-EQ(AA) contents higher than the corresponding values obtained with jar cultures. Under stationary cultivation, G. sordulenta produced approximately 50% higher content in both RS-EQ(AA) and RP-EQ(AA) than the medicinal mushroom G. frondosa. Phenolics correlated with RS-EQ(AA) and RP-EQ(AA) in G. gargal and with RP-EQ(AA) in G. sordulenta; besides, thin-layer chromatography showed these compounds to be at least in part related to the RS capacity. It is concluded that G. gargal and G. sordulenta mycelia are excellent sources of antioxidant metabolites.

An in vitro high‐throughput assay for screening reproductive and toxic effects of anticancer compounds

An in vitro assay was developed that simultaneously tested the effects of anticancer drug candidates on cytotoxicity, hormone synthesis, and gonadotrophin responsiveness using the choriocarcinoma JAr cell line. JAr culture conditions were optimized and then cells were exposed to a marine mollusc extract in the presence and absence of hCG. The intra- and interassay coefficients of variation of the optimized 1 H thiazolyl blue tetrazolium bromide assay were 11.3% and 10.9%, respectively. hCG (1,000 mIU/mL) increased progesterone (P4) synthesis after 24 H (P<0.05). The mollusc extract significantly decreased cell viability, with the IC50 affected by incubation time, but not hCG. P4 synthesis was inhibited at low concentrations of the anticancer extract, but stimulated at the highest concentration, and complex interactions of P4 were also found with hCG. In conclusion, the optimized assay is useful to characterize the effects of novel drugs on cytotoxicity, basal, and gonadotrophin-stimulated P4 synthesis in vitro, and can be used to inform subsequent in vivo studies.

Interleukin-12 augments cytolytic activity of peripheral and decidual lymphocytes against choriocarcinoma cell lines and primary culture human placental trophoblasts.

Human trophoblasts are tolerant to the maternal immune system, but susceptible to interleukin (IL)-2-activated lymphocytes. IL-12 is also a key cytokine in the induction of cytotoxic responses. We administered IL-12 to peripheral blood lymphocytes (PBLs) and to decidual lymphocytes (DLs) and studied resulting cytotoxicity against trophoblasts. PBLs and DLs were stimulated with rIL-2 and/or rIL-12 for 48 hr in vitro. Cytotoxicity against the choriocarcinoma cell line JEG-3, JAR, and primary culture trophoblasts were examined by LDH release assay. The proliferative response was estimated by MTT assay. Expression of cytotoxic factors was studied by reverse transcriptase-polymerase chain reaction (RT-PCR). Whereas IL-12 alone produced a modest enhancement in cytotoxicity of PBLs and DLs, the combination of IL-2 and IL-12 was most effective in trophoblast cell lysis. IL-12 enhanced the mRNA expression of T-cell specific serine protease (TSP, granzyme B) and FasL in DLs, but the expression of perforin was unchanged. Expression of these cytotoxic factors in PBLs was up-regulated by IL-12. Our findings indicate critical roles of IL-12 in the activation of maternal lymphocytes, which could possibly result in pregnancy failure syndromes.

Nodulation Interaction of the Soybean Allele Rj2 with Asiatic Isolates of Bradyrhizobium japonicum

The dominant allele Rj2 in soybean, Glycine max (L.) Merr., restricts nodulation with Bradyrhizobium japonicum (Jordan) strains USDA 7, USDA 14, and USDA 122, while the recessive allele rj2 permits normal nodulation. To determine the frequency of nodulation restriction by the Rj2 allele of soybean, 89 bradyrhizobial isolates from Asia were tested in Leonard jar culture for nodulation response with near isogenic plant lines differing at the Rj2 locus. Fifty‐one percent of the isolates tested elicited the nodulation restriction of the Rj2 allele. No geographic pattern in the occurrence of Rj2‐restricted strains was discerned. Nearly half (47%) of the isolates restricted by the Rj2 allele were from Serogroup 6. Twenty‐one of 25 Serogroup 6 strains were restricted by the Rj2 allele. Since some strains in Serogroup 6 were not restricted by the Rj2 allele and some strains not in Serogroup 6 were restricted, specific serological reaction per se is not essential or necessary for eliciting the action of the Rj2 allele. It appears that the Rj2 rejection is not as related to serotype as had been assumed.

The Rj4 allele in soybean represses nodulation by chlorosis-inducing bradyrhizobia classified as DNA homology group II by antibiotic resistance profiles

To determine the relationship between nodulation restriction by the Rj4 allele of soybean, rhizobitoxine-induced chlorosis, and taxonomic grouping of bradyrhizobia, 119 bradyrhizobial isolates were tested in Leonard jar culture for nodulation response and chlorosis induction. In addition to strain USDA 61, the strain originally reported as defining the Rj4 response, eight other isolates (i.e., USDA 62, 83, 94, 238, 252, 259, 260, and 340) were discovered to elicit the nodulation interdiction of the Rj4 allele. Only 16% of all the bradyrhizobial strains tested induced chlorosis, but seven of the nine strains (78%) interdicted by the Rj4 allele were chlorosis-inducing strains. Furthermore, in tests for antibiotic resistance profile, eight of the nine interdicted strains (89%) were classed in DNA homology group II. This evidence suggests that the Rj4 allele has a positive value to the host plant in shielding it from nodulation by certain chlorosis-inducing bradyrhizobia of a DNA homology group with impaired efficiency of nitrogen fixation with soybean.

O-Glycosylation of the α-Subunit Does not Limit the Assembly of Chorionic Gonadotropin αβ Dimer in Human Malignant and Nonmalignant Trophoblast Cells*

Biosynthetic experiments were carried out in cultures of human malignant trophoblast cells (the JAR cell line) and in explants of normal first trimester human placental tissue to test the hypothesis that the O-glycosylation of the glycoprotein hormone alpha-subunit at Thr-39 regulates the assembly of the CG alpha beta dimer. This modification of alpha has been shown to ablate its ability to combine in vitro with the beta-subunit of bovine LH and might explain why JAR cells and placental explants secrete uncombined alpha- and beta-subunits in addition to the hCG alpha beta dimer. We have previously detected an O-linked carbohydrate chain at Thr-39 in preparations of secreted free alpha-subunit, but not dimer CG alpha from JAR culture medium. We report here evidence that the O-glycosylation of alpha does not regulate the biosynthetic assembly of the hCG dimer in cultures of JAR choriocarcinoma cells or first trimester placental explants. The intracellular precursor forms of alpha and beta that accumulate in the endoplasmic reticulum and combine in that compartment are not yet modified with O-linked carbohydrate, as determined by measurements of their [3H]galactosamine content after biosynthetic labeling of amino sugars with [3H]glucosamine. Furthermore, only half of the free alpha-subunit secreted by JAR cells and less than 10% of free alpha secreted by 10-week-old placental explants received the O-linked chain. This was shown by determining the ratio of the unglycosylated and glycosylated forms of the tryptic peptide from free alpha that contains the O-glycosylation site (residues 36-42). Based on these findings, we make the following conclusions. 1) O-Glycosylation of alpha-subunit is a late event in the secretory pathway of trophoblasts compared to the rapid combination in the rough endoplasmic reticulum of hCG subunit precursors to form alpha beta dimer. 2) Association of alpha with beta precludes the subsequent addition of the glycan to alpha at Thr-39. 3) The alpha molecules that fail to combine with beta in the endoplasmic reticulum are substrates for the later addition of O-linked carbohydrate, presumably in the Golgi complex, but only a fraction of the free alpha molecules are modified with O-linked carbohydrate.

Biosynthesis and Deposition of a Noncovalent Laminin-Heparan Sulfate Proteoglycan Complex and Other Basal Lamina Components by a Human Malignant Cell Line

The basal lamina components laminin, heparan sulfate proteoglycan (HSPG), and type IV collagen were synthesized and codeposited in the extracellular matrix (ECM) by a cultured human cell line from gestational choriocarcinoma (JAR). Laminin and HSPG formed a noncovalent complex detected by the coimmunoprecipitation of HSPG with laminin from cell lysates and culture media. The complex was stable in the cell lysis buffer that contained detergents (1% Triton X-100, 0.5% deoxycholate, and 0.1% sodium dodecyl sulfate) and sodium chloride (from 0.15 to 1.0 M), but was dissociated by adding 8 M urea to the detergent lysates. Even though JAR cells produced roughly equal amounts of HSPG and chondroitin sulfate proteoglycan, only HSPG complexed with laminin, suggesting a specific interaction between these basal lamina components. The laminin-HSPG complex was deposited and retained in the ECM. This was shown biochemically by isolating an enriched fraction of ECM from JAR cells cultured on native type I collagen gels. At steady state, more than half (52%) of the laminin-HSPG in the culture was recovered in the ECM fraction, in contrast to 16% of the total laminin and 29% of the total type IV collagen, which were secreted to a greater extent than laminin-HSPG into the culture medium. The retention of the laminin-HSPG complex in the ECM suggests that it may participate in the assembly of the basal lamina-like extracellular matrix deposited by JAR cultures. Omission of ascorbate from the culture medium abolished the ECM deposition of type IV collagen but had little effect on the deposition of laminin or laminin-HSPG. This demonstrates that the stable deposition of laminin-HSPG and laminin in the collagen-based choriocarcinoma cultures is not dependent on an assembled network of type IV collagen.

Gametocytogenesis induction in cultured Plasmodium falciparum and further development of the gametocytes to ookinetes in prolonged culture.

Gametocytogenesis induction medium (RPMI-FSC reactive medium), prepared with a mixture of the culture supernatant from anti-Plasmodium falciparum antibody-producing hybridoma cells and hybridoma cell lysate, consistently induced gametocytogenesis in cultured P. falciparum. Gametocytogenesis was induced in three strains (0662, FCB 1, and FCR-3) and one clone (R.FCR-3) as has previously been described in two other strains (Ono et al. 1986). Both caffeine (2 mM/ml) and calmodulin (5 micrograms/ml) enhanced the effects of RPMI-FSC reactive medium. Furthermore, gametocytes that appeared after treatment with RPMI-FSC reactive medium continued their development to ookinete formation in candle jar culture with Waymouth's medium.

Wise Anaerobic Work Station: an evaluation.

The Wise Anaerobic Work Station permitted the growth of type cultures of a range of fastidious obligate anaerobes. In the isolation of anaerobic bacteria from clinical specimens the performance equalled that of a standard anaerobic jar incubated for 48 hours, and gave results superior to those obtained when incubation of jar cultures was interrupted at 24 hours. The Wise Anaerobic Work Station was easy to use and provided comfortable working conditions; running largely on industrial nitrogen, it was more economic than anaerobic cabinets or jars using conventional gas mixtures.

ニオイテンジクアオイ不定芽組織のジャーファーメンター培養

We were able to develop a shoot-forming culture of Pelargonium graveolens in jar fermentors by a modification of Linsmaier-Skoog's medium with 10-5M indoleacetic acid and 10-6M 6-benzyl-adenine. Young adventitious shoots (10g fresh weight) which had been cultured on agar media were inoculated into 5-liter jar fermentors containing 3-liters of medium and cultured under continuous illumination at 25°C with agitation of 3 liters/min (1vvm) aeration through sintered glassfilter. After 3 weeks, the yield was 145g fresh weight. The GC-pattern of volatile compounds in the extract of the shoot-forming culture was similar to that of natural Pelargonium leaf oil, and these volatile compounds in the shoot-forming culture were identified with standard compounds. But the content of citronellol (a major constituent of natural Pelargonium leaf oil) in the culture was less than that in intact leaves. With the use of 10-5M indolebutyric acid as auxin and reduced concentration of NH4+, the weight of the shoot-forming culture increased to 37.5-fold during 3 weeks of jar culture.

Nitrogen metabolism and chloramphenicol production in Streptomyces venezuelae.

The relationship between chloramphenicol production and nitrogen metabolism in Streptomyces venezuelae was examined in stirred jar cultures under pH control. Nitrogen sources that supported rapid biomass accumulation gave low rates of antibiotic synthesis during growth. This was consistent with a general incompatibility between fast growth and high yields of chloramphenicol. In media where the growth rate was reduced below the attainable maximum by the rate at which nitrogen could be assimilated, chloramphenicol production was associated with biomass accumulation. Enzymes that are potentially associated with nitrogen assimilation pathways were assayed in cultures supplied with nitrogen sources supporting markedly different growth rates. The results indicated that glutamine synthetase and alanine dehydrogenase levels were relatively insensitive to changes in growth rate and nitrogen source depletion. Glutamate dehydrogenase and glutamate synthase, on the other hand, showed high activity in cultures assimilating ammonium nitrogen and markedly decreased activity with poorer nitrogen sources or when ammonium was depleted. If chloramphenicol biosynthesis is coordinately controlled by mechanisms that regulate nitrogen assimilation, glutamate synthase and glutamate dehydrogenase are the most likely enzymes that manifest the regulatory linkage.

Nodule‐Like Structres Induced on Peanut by Chlorosis Producing Strains of Rhizobium Classified as R. Japonicum1

Some strains of rhizobia produce a rhizobitoxine‐induced foliar chlorosis on soybean, Glycine max (L.) Merr., and have a limited ability to nodulate the nodulation restrictive genotype (rj1). This study was undertaken to determine and compare the effect of chlorosisinducing or rj1‐compatible strains and nonchlorosis‐inducing strains on another legume, peanut (Arachis hypogaea L., cultivars ‘Starr’ and ‘Spancross’), in Leonard jar culture. Ten different rhizobial strains that induce chlorosis or nodulate rj1‐plants, produced swellings at the nodulation sites on peanut roots. No rhizobial‐induced chlorosis symptoms developed on either peanut cultivar. Control peanut plants inoculated with six different nonchlorosis‐inducing strains of Rhizobium japonicum (Kirchner) Buchanan did not have swellings. The clustering of three distinctive characteristics, a) ability to nodule rj1 plants, b) the propensity to produce rhizobitoxine‐induced chlorosis symptoms on soybean, and c) the ability to induce nodule‐like swellings on peanut roots, in a common population of rhizobia suggests that these rhizobia may represent a taxonomically distinct group. These strains appear defective in their symbioses with both soybean and peanut, suggesting that they may have a symbiotic homology for a legume host other than either soybean or peanut.

Lactic acid production from 1,2-propanediol by jar culture and resting cells of Arthrobacter oxydans.

Arthrobacter oxydans (PG21–1) which was isolated from soil preferably assimilated 1,2-propanediol and produced remarkable amount of lactic acid. Cultural conditions for lactic acid production from 1,2-propanediol were studied using a 15 liter jar fermentor. Optimum pH and urea concentration for the production of lactic acid were 8.0 and 0.3%, respectively. Under these conditions, the yield of lactic acid was 17.8 g/liter in 7 day cultivation. About 70 % of the utilized 1,2-propanediol was converted to lactic acid. The production of lactic acid by resting cells was also investigated. Resting cells of Arthrobacter oxydans showed a high activity to produce lactic acid at alkaline pH and the amount of the acid reached to 10.2 g/liter in 12 hr incubation. The accumulation of lactic acid at alkaline side was considered to due to the strong suppression of the degradation of lactic acid.

A sterile sea water circulating system for the jar culture of developing fish eggs

A sterile sea water circulating system for the jar culture of developing fish eggs is described. An ultra–violet filter unit is used to sterilise the water and the system is one of a double‐circulation the advantages of which are discussed. This is an effective method for maintaining the developing embryos of Blennius pholis L. under controlled conditions, and would be applicable to studies on other demersal fish species or marine organisms.

Modifications for the Jar Culture of Trout Eggs

Modifications for the Jar Culture of Trout Eggs

Jar Culture of Trout Eggs

Jar Culture of Trout Eggs