Cryopreservation of Sheep Red Blood Cells

The protection of sheep erythrocytes at freezing temperatures was investigated using glycerol, dimethylsulfoxide (DMSO), glucose and four different types of polyvinylpyrrolidone (PVP) as cryoprotective agents. Depending on type (molecular weight) and concentration good protection was obtained with PVP, whereas glycerol, DMSO and glucose were unsatisfactory. Recovery of cells after thawing was most successful when the cells had been frozen at a concentration of 1–2 × 109 cells/ml. No cells tolerated freezing at −20 °G. Best results were obtained when the cells were frozen directly in liquid nitrogen (−196°G).     

Influences of the priming procedure and saline circulation conditions on polyvinylpyrrolidone in vitro elution from polysulfone membrane dialyzers

In hemodialysis (HD), the patient's blood is purified via circulation in an extracorporeal circuit containing a dialyzer. In the manufacturing process of polysulfone (PSu) membrane dialyzers, the membranes are hydrophilized via the addition of the hydrophilic agent polyvinylpyrrolidone (PVP) to increase their hydraulic permeability. The elution of PVP from the membrane reduces the membrane's hydraulic permeability, and the eluted PVP could cause adverse effects in the human body. Therefore, it is important to identify the factors that induce PVP elution from PSu dialyzer membranes to improve the efficiency and safety of HD. In the present study, experimental circuits connecting each of the three types of PSu membrane dialyzers that had been sterilized, using gamma irradiation, autoclaving, or in-line steam methods, were prepared. After the dialyzers were primed, saline was circulated in the circuits at a flow rate of 100 mL/min or 200 mL/min. At 0, 2, 4, 6, and 8 h after circulation was initiated, the amount of PVP eluted from the PSu membranes in vitro was determined. In this experimental setting, longer the circulation duration, greater the amount of PVP eluted from the PSu membranes of the tested dialyzers; however, the flow rate did not influence the in vitro elution of PVP. Furthermore, the immersion of the dialyzer membranes in saline for 24 h strongly facilitated the in vitro elution of PVP. In sum, these results suggest that the duration of PSu membrane incubation in saline is a determinant of the level of PVP elution from the PSu membrane dialyzers.     

The mediator of cellular immunity: XI. Origin and development of MIF producing lymphocytes

Thoracic duct lymphocytes obtained from rats infected with Listeria monocytogenes were characterized with respect to size, turnover and their capacity to release macrophage migration inhibitory factor (MIF). Cells responsive to Listerial antigens (LMA) in the MIF assay were identified in lymph during the first week of an immunizing infection. These were immunoblasts or large lymphocytes, as evidenced by their sedimentation with S phase lymphocytes at unit gravity. When labeled cells from the lymph of Listeria-infected donors were infused into similarly infected recipients, donor S phase lymphocytes localized rapidly, and in substantial numbers, in peritoneal exudates induced by the unrelated organism, F. tularensis. Within this immigrant population were cells which conferred immunity against L. monocytogenes and released MIF in cultures containing LMA. Exudates harvested 36 hr or 61 hr after stimulation contained labeled lymphocytes that were smaller than the S phase cells recovered during the early post-induction period. The observed shift of radioactivity from large to smaller lymphocytes was parallelled by a shift MIF production to exudate fractions containing smaller cells. The MIF producing cells in exudates of advancing age also exhibited increasing resistance to inhibition by vinblastine. These findings suggest that MIF is released by a family of lymphocytes—large, medium and small. LMA-responsive lymphocytes are delivered to the thoracic duct soon after their formation, at a stage in development when they can be stimulated to release only low levels of MIF. These mediator producing cells circulate briefly in the blood and differentiate fully only after they extravasate into inflammatory foci.     

Effect of lectins on the interactions between human peripheral blood lymphocytes and monocytes

Interactions between normal human peripheral blood T lymphocytes and monocytes were investigated by measuring the in vitro cellular adherence of these cells in the presence and in the absence of mitogens. Concanavalin A (Con A), lentil lectin (Lc), and phytohemagglutinin (PHA) in mitogenic doses increased 15 to 20 times the binding of T lymphocytes to monocytes. The lectin-induced binding was similar to that produced by neuraminidase-gal-actose-oxidase treatment. A good correlation was found between the early cellular adherence induced by these lectins and by neuraminidase-galactose-oxidase and the blastogenesis of the T lymphocytes measured after 3 days of culture by [³H]thymidine uptake. However, wheat germ agglutinin (WGA), a nonmitogenic lectin, also increased the binding of T lymphocytes to monocytes. Addition of specific carbohydrates completely inhibited the cellular interactions induced by lectins. Peanut agglutinin (PNA) induced adherence of lymphocytes only after treatment of these cells with neuraminidase. Striking differences were not found between the lectin-induced adherence observed with autologous and heterologous cells. Killing of monocytes abolished entirely the lectin-induced adherence of lymphocytes, however killed T lymphocytes were still able to interact weakly with live monocytes. Dexamethasone was found to be a potent inhibitor of mitogen-induced cellular interactions.     

In vitro glucocorticoid inhibition of steroid-treated residual circulatory human lymphocytes

Various doses of glucocorticoids given in vivo caused a similar degree of maximal lymphopenia. The sensitivities of mitogen-induced proliferation to the suppressive effects of glucocorticoid added in vitro were studied in residual lymphocytes obtained after steroid injection. Methylprednisolone (MP) administered intravenously depleted circulatory lymphocytes and reduced markedly the proliferative responses of residual cells to mitogens (phytohemagglutinin and concanavalin A) 4 to 8 hr after the injection. The addition of MP in vitro to the residual cells further inhibited the cell proliferation. The degrees of proliferation inhibition induced by in vitro MP were compared in cells obtained at various intervals after MP injection. At each specific mitogen concentration, lymphocytes obtained at various intervals were inhibited to a similar degree by MP in the cultures. There was no evidence that cells obtained at the period of maximal lymphopenia, 4 to 8 hr after MP injection, were more resistant to the inhibition of glucocorticoid added in vitro. Hence, the residual lymphocytes were not “steroid-resistant” in the sense of proliferative responses to T-cell mitogens. These results indicate the mechanism of lymphocyte sequestration is unrelated to the suppressive effects of glucocorticoid on cell proliferation.     

The influence of phase shift in the light-dark cycle on humoral immune responses of mice to sheep red blood cells and polyvinylpyrrolidone

The influence of a phase shift in the light-dark cycle on humoral immune responses against sheep red blood cells (SRBC), a thymus-dependent antigen, and against polyvinylpyrrolidone (PVP), a thymus-independent antigen, was studied by using 180 BALB/c mice and 150 C3H/HeN mice. Significant suppression of the immune response to SRBC and the number of splenic plaque-forming cells (PFC) and hemagglutination (HA) titers was observed on days 5 and 6 after inversion of the light-dark cycle. On the other hand, the number of splenic PFC and HA titers in the blood against PVP were minimally suppressed by the phase shift in C3H/HeN mice, except for distortion of the rhythmicity. Corticosterone levels in the blood on days 5 and 6 after inversion were higher than those under a normal lighting regimen. The appearance of the high corticosterone level in the blood after the inversion almost concurred with the suppression of the immune response to SRBC. A decrease of the proportion of splenic T cells was also observed on day 6 after the inversion. These results show that a phase shift in the light-dark cycle provokes suppression of the immune response to SRBC, possibly through an increase of secretion of corticosterone after light-dark inversion, which induces a decrease of the proportion of T lymphocytes in the spleen.     

Cryoprotection of mammalian cells in tissue culture with polymers; possible mechanisms

Studies were undertaken to more clearly define the mechanism of cryoprotection by polymers. Significant cryoprotection of Chinese hamster cells in tissue culture was found in the presence of hydroxyethyl starch (HES), polyvinylpyrrolidone (PVP), and dextran. The addition of PVP to the medium after thawing did not increase the survival of these cells. The presence of PVP in the medium was shown to have no effect on the transport mechanism for alanine in unfrozen cells. The source of freeze-thaw injury did not appear to be due to a direct effect on this transport mechanism. Several physical parameters of polymeric solutions were monitored at subzero temperatures. The freezing point depression was found to increase dramatically at higher polymer concentrations. Tests on the NaCl concentration in the liquid fraction of partially frozen solutions showed that the increase in salt concentration with decreasing temperature was similar in the presence of 10% PVP or 2.5% DMSO, two agents which gave similar cryoprotection at these concentrations. NMR studies showed that polymers could retain water in the liquid state at temperatures as low as −35° C, and that the remaining water was highly structured. The cryoprotective properties of polymers appear to reside in their ability to alter the physical properties of solutions during the freezing process rather than in direct effects on cell membranes.     

In vivo and in vitro ethylene oxide exposure of human lymphocytes assessed by chemical stimulation of unscheduled DNA synthesis

Factory workers exposed to ethylene oxide (EO), 0.5–1.0 ppm in factory air, together with matched controls from the same factory, were examined for evidence of toxic exposure by measurement of unscheduled DNA synthesis (UDS) induced by N-acetoxy-2-acetylaminofluorene (NA-AAF) and of chromosome aberrations in peripheral lymphocytes.The total chromatid gaps plus breaks were significantly elevated and NA-AAF-induced UDS was significantly reduced in the EO-exposed group as compared with the unexposed control group. The NA-AAF-induced UDS values negatively correlated to the duration (yr) of EO exposure (r = ?0.45, p < 0.02) and the number of chromosome breaks (r = ?0.61, p < 0.05), indicating an inhibition in vivo of DNA-repair capacity by EO. These data were verified in vitro by biochemical and autoradiographic studies of EO-induced UDS in human blood cells. Above 2 mM EO, UDS was inhibited in lymphocytes whether they were cultured for 24 or 122 h after alkylation with EO. Even at the subtoxic EO dose of 0.1 mM, lymphocytes were sensitized to additional exposures of NA-AAF, so that cytotoxicity was increased to 40% compared with 5% for the controls even though UDS was unaffected.It is concluded that EO was toxic to lymphocytes, even when they were sensitized at non-toxic EO doses to the cytotoxic action of other mutagens (e.g. NA-AAF), and the cells that did survive above 2 mM EO were inhibited in their DNA-repair capacity as judged by reduced UDS.     

Synthesis and Characterization of Pectin/PVP Hydrogel Membranes for Drug Delivery System

The purpose of the present study was to develop and design pectin and polyvinyl pyrrolidone (PVP) blended hydrogel membranes (PEVP), with different pectin: PVP ratios (1:0.2, 1:0.4, 1:0.6, 1:0.8 and 1:1 w/w), which were prepared by using a conventional solution casting technique. An attempt has been made to characterize the hydrogel membranes by various instrumental techniques like, FTIR (Fourier transform infrared) spectroscopy, X-ray diffraction (XRD), Differential scanning calorimetry (DSC), tensile strength test and scanning electron microscopy (SEM). The release patterns of the drug (salicylic acid) from the hydrogel membrane were done in three different release mediums (pH 1.4, pH 7.4 and distilled water) and samples were analyzed spectrophotometrically at 294 nm wavelength on a UV Vis spectrophotometer. MTT assay was done to ensure cytocompatibility of the pectin/PVP hydrogel membranes using B16 melanoma cells. FTIR spectroscopy indicated the presence of secondary amide (I) absorption bands. The XRD study shows decrease in crystallinity of the hydrogel membranes with increase in PVP ratio. DSC study shows an increase in T _g of pectin after blending with PVP. It was found that tensile strength increases with increasing PVP ratios in the hydrogel membranes. The prepared hydrogel membranes were found to be biocompatible with B16 melanoma cells.     

Some problems of spontaneous and induced mutagenesis in mammals and man.

The culture time of rabbit lymphocytes (41–42 h) that provides cells in their first post-stimulation mitosis, was estimated on the basis of the mitotic index, dicentric yield and presence of the cells with these aberrations unaccompanied by acentric fragments, studied as a function of culture duration. The cells obtained in metaphase from cultures terminated at this time displayed no donor-to-donor variation where induction of dicentrics by X-rays was concerned.Rabbit venous blood was irradiated in vitro with a range of X- and gamma-ray doses, and dose-effect curves were obtained by regression analysis. Sixteen rabbits were irradiated in vivo (uniform whole-body irradiation), and blood was sampled 10 min, 6, 24, and 48 h after exposure. The frequency of dicentrics in the lymphocytes cultured did not change significantly over the first 24 h after irradiation. Dose-effect relationships in vivo fell within one standard error confidence limits of the respective curves in vitro. The authors conclude that the latter may be used for estimation of dose in vivo under conditions of homogeneous whole-body irradiation.     

Studies on the recognition of xenogeneic cells by nonimmune macrophages. I. Destruction of chicken fibroblasts in vitro by murine macrophages.

Human cord blood lymphocytes were compared with adult lymphocytes with regard to proportions of cells with surface markers for surface immunoglobulin (Ig), receptors for C′3 and the Fc-portion of IgG, as well as two types of erythrocyte rosettes (rapid and late E-rosettes). A significant decrease (P < 0.02 ? 0.05) in both early and late E-rosettes was noted when cord cells were compared to adult lymphocytes. After 20 hr of incubation at 37 °C, proportions of cells bearing Fc receptors in cord blood samples showed striking increments (P < 0.001) when compared with adult lymphocytes. T cell enrichment studies and sequential depletion of cells bearing Fc receptors as well as E-rosette forming cells indicated that the precursors of cells generating Fc receptors in vitro did not arise from cells with Fc receptors or T cell markers.     

The effects of corticosteroids on immunoregulation in sarcoidosis.

The effects of in vivo hydrocortisone administration on the kinetics and functional capabilities of cells involved in the immune response in sarcoidosis were examined. Untreated sarcoidosis patients have a decrease in the absolute numbers of circulating T lymphocytes (P < 0.05). However, with regard to the proportions of T lymphocyte subpopulations, there is an increase in the relative proportions of IgG Fc receptor positive T cells (T_G) (P < 0.01), which have suppressor capabilities in certain in vitro systems of mitogen-induced antibody production, and a relative decrease in IgM Fc receptor positive T lymphocytes (T^M) which have helper effects in this system (P < 0.05). Additionally, sarcoidosis patients have circulating “suppressor” monocytes capable of suppressing anti-sheep red blood cell (SRBC) plaque-forming cell (PFC) responses by pokeweed mitogen (PWM)-stimulated lymphocytes. The in vitro removal of this cell abrogated this depressed response (P < 0.01). Intravenous administration of hydrocortisone produced a transient absolute T lymphocytopenia (P < 0.01) accompanied by a relative increase in T^G cells (P < 0.01) and a relative decrease in T_M cells (P < 0.02). Four hours after hydrocortisone therapy, at the point of maximal hydrocortisone-induced monocytopenia (P < 0.01), the suppressed ability of sarcoidosis lymphocytes to synthesize and secrete in vitro anti-SRBC antibody after polyclonal activation was corrected (P < 0.01), and PFC responses comparable to those seen in untreated normal subjects were obtained. These studies demonstrate that corticosteroid administration has profound effects on certain in vitro demonstrable immunoregulatory abnormalities in sarcoidosis.     

Induction of interferon production in mouse spleen cell cultures by corynebacterium parvum.

Spleen cells of CS7BL/6 mice produced considerable amounts of interferon (IF) in vitro when tested 5 to 20 days after injection of killed Corynebacterium parvum. Interferon was also produced when C. parvum was added in vitro to spleen cell cultures of previously untreated mice. High levels were detected after 1 day of culture with some increment during subsequent days. In a number of experiments IF was also produced in untreated control cultures but only after prolonged cultivation and not after 1 day. The highest levels of IF were usually obtained when spleen cells of C. parvum-treated mice were challenged with additional C. parvum in vitro. The IF induced by C. parvum shared certain physicochemical properties with a tested immune IF and was not neutralized by an antiserum raised against a type I IF. Spleen cells of nu/nu mice and spleen cells treated by anti-θ serum plus complement did not differ from their respective controls, indicating that production of IF did not require mature T lymphocytes. Removal of B lymphocytes by nylon wool columns abolished the capacity of spleen cells to produce IF. When spleen cells were freed of adherent cells by the use of plastic surfaces, they no longer produced IF. Peritoneal exudate macrophages (PEC), which by themselves did not produce IF, in small numbers reconstituted nonadherent spleen cells. Nylon column-treated spleen cells, however, could not be restored by PEC. It is concluded that IF upon challenge with C. parvum is produced by B lymphocytes and requires the help of macrophages.     

Evaluation of genotoxicity, cytotoxicity and cytostasis in human lymphocytes exposed to patulin by using the cytokinesis-block micronucleus cytome (CBMN cyt) assay

Patulin (PAT) is a fungal secondary metabolite commonly present in apples and apple products. In the present study, PAT was evaluated for its genotoxic, cytotoxic and cytostatic effects to human peripheral blood lymphocytes by using the cytokinesis-block micronucleus cytome (CBMN Cyt) assay. Lymphocyte cultures were treated with PAT at the following concentrations, 0.1, 0.3, 0.5, 1.0, 2.5, 5.0, and 7.5 μM, as well as 0.5 μM mitomycin c (MMC) as a positive control and dimethyl sulfoxide (DMSO) as a vehicle control. PAT was found to induce nucleoplasmic bridges (NPBs) at 5.0 and 7.5 μM concentrations (P?<?0.05), apoptotic cells at 0.1, 1.0, 5.0 μM (P?<?0.05), 7.5 μM concentrations (P?<?0.01) and necrotic cells at 0.3 and 2.5 μM (P?<?0.05), 5.0 and 7.5 μM (P?<?0.01) concentrations in human lymphocytes. The 2.5, 5.0, and 7.5 μM PAT concentrations also led to a clear decrease in the nuclear division index (NDI) (P?<?0.05). PAT caused a significant dose-dependent increase in the number cells of NPBs, in the frequency of apoptotic and necrotic cells, and a significant dose-dependent decrease in the NDI values in lymphocytes. These results indicate that PAT at high concentrations is genotoxic, cytotoxic and cytostatic in cultured human lymphocytes.     

Role of lymphocyte surface determinants in lymph node homing

Thoracic duct lymphocytes briefly incubated in vitro with trypsin and then infused into syngeneic rats are unable to migrate into lymph nodes. Trypsin-treated lymphocytes incubated at 37 °C in the absence of enzyme for 12 hr recovered their lymph node homing properties. In vitro recovery did not occur if the cells were cultured at 17 °C. Evidence was obtained that trypsin cleaved sialyglycoproteins from the surface of lymphocytes and that these determinants reappeared after the cells were maintained at 37 °C for 24 hr.Puromycin added to cultures of normal lymphocytes for 3 hr before infusion markedly reduced the recovery of donor cells in lymph nodes. The results suggest that surface determinants of recirculating lymphocytes essential for homing into lymph nodes may be rapidly turned over.     

Selective T cell killing of human lymphocytes by ultraviolet radiation

The effects of ultraviolet radiation (uv) on human B and T lymphocytes were studied. In vitro studies showed that T lymphocytes were more sensitive to uv than B lymphocytes as assessed by eosin-dye exclusion. Following uv exposure, the viable lymphocytes responded to mitogens (PHA, PWM), and functional B lymphocytes were present at a time when no viable T cells were detected. Varying doses of uv were required to abrogate different in vitro responses (proliferative response to antigen or allogeneic cells, MIF production, and cell-mediated lympholysis). In vivo, uv was able to diminish an established cutaneous delayed hypersensitivity response. In vitro uv treatment of parental mouse spleen cells eliminated a graft-versus-host reaction in F₁ recipients as determined by the spleen index. The basis for the differential effect of uv on B and T lymphocyte viability and functional responses is unknown.     

Altered expression of inflammatory cytokine receptors in response to LPS challenge through interaction between intestinal epithelial cells and lymphocytes of Peyer's patch

The intense innate immunological activities occurring at the enteric mucosal surface involve interactions between intestinal epithelial cells and immune cells. Our previous studies have indicated that Peyer's patch lymphocytes may modulate intestinal epithelial barrier and ion transport function in homeostasis and host defense via cell-cell contact as well as cytokine signaling. The present study was undertaken using the established co-culture system of Caco-2 epithelial cells with lymphocytes of Peyer's patch to investigate the expression of IL-8 and IL-6 cytokines and cytokine receptors in the co-culture system after challenge with Shigella F2a-12 lipopolysaccharide (LPS). The human colonic epithelial cell line Caco-2 was co-cultured with freshly isolated lymphocytes from the murine Peyer's patch either in the mixed or separated (isolated but permeable compartments) co-culture configuration, and was challenged with Shigella F2a-12 LPS for 8 h. The level of mRNA expressions of human interleukin-8 (hIL-8), human interleukin-8 receptor (hIL-8R), mouse interleukin-8 receptor (mIL-8R), mouse interleukin-6 (mIL-6), mouse interleukin-6 receptor (mIL-6R) and human interleukin-6 receptor (hIL-6R) was examined by semi-quantitative PCR. In both co-culture groups, hIL-8 expression of Caco-2 cells was decreased, and hIL-8R expression was increased compared to the Caco-2 alone group. Upon LPS challenge, hIL-8 expression from the Caco-2 cells of both co-culture groups was higher than in the Caco-2 control group. The increased hIL-8 expression of Caco-2 cells in the separated co-culture group is correlated with a decreased hIL-8R expression and an increased mIL-8R expression. In the mixed co-culture group, the increased expression of hIL-8 was associated with the upregulated hIL-8R expression on Caco-2 cells and downregulated mIL-8R on murine Peyer's patch lymphocytes (PPL). mIL-6 expression from mouse PPL was also upregulated by LPS in mixed co-culture. However, upon the treatment with LPS, hIL-6R expression of Caco-2 cells was decreased in the mixed co-culture, but increased in separated co-culture. The data suggest that release of hIL-8 from epithelial cells may act on lymphocytes through a paracrine pathway, but it may also act on the epithelial cells themselves via an autocrine pathway. The data also suggest that the release of mIL-6 from Peyer's patch lymphocytes affects epithelial cells in a paracrine fashion.     

Characterization of subpopulations of T lymphocytes: I. Separation and functional studies of peripheral T-cells binding different amounts of fluorescent anti-Thy 1.2 (theta) antibody using a fluorescence-activated cell sorter (FACS)

Peripheral T lymphocytes in mice can be distinguished by the presence of the Thy 1.2 (theta) cell surface antigen. The fluorescence-activated cell sorter (FACS) was used to analyze and separate T-cells from peripheral lymphoid cell suspensions after incubation with fluorescein-labeled anti-Thy 1.2 (F anti-Thy 1.2). Stained cells were markedly reduced in nu/nu mice, in mice carrying the Thy 1.1 allele (theta-AKR), and were not seen after incubation with anti-Thy 1.2 that had been absorbed with CBA brain. According to these criteria, the stained cells were termed “T lymphocytes.”Among the T lymphocytes, there was considerable heterogeneity of fluorescent staining. The FACS was used to separate T-cells from other cells and further to separate T-cells with high intensity F anti-Thy 1.2 fluorescence (bright T-cells) from those with less F anti-Thy 1.2 fluorescence (dull T-cells). Separated bright T and dull T lymphocytes were shown to have several different functional properties. Dull T-cells appeared more sensitive to small doses of ALS in vivo, homed to lymph node in higher proportions than did bright T-cells, and were not affected by the short-term effects of thymectomy in adult life. Bright T lymphocytes, by contrast, were resistant to the in vivo effects of ALS, homed preferentially to spleen rather than lymph node in irradiated hosts, and were reduced shortly after adult thymectomy. Separated populations of bright and dull T-cells showed reduced ability to produce cytotoxic activity after in vitro sensitization, while mixtures of these two subpopulations of T-cells produced synergistic cytototoxic responses. The ontogenic and functional implications of these findings are discussed.     

Induction of unscheduled DNA synthesis by the carcinogen 7-bromomethylbenz(a)anthracene and its removal from the DNA of normal and xeroderma pigmentosum lymphocytes

The carcinogen 7-bromomethylbenz(a)anthracene (BBA), which can bind strongly to DNA, induces unscheduled DNA synthesis (DNA repair) in normal lymphocytes but almost none in lymphocytes from patients with Xeroderma pigmentosum (XP), and inherited disease known to be defective in excision repair of ultraviolet-damaged DNA. We studied [³H]BBA's ability to bind to DNA of normal and XP lymphocytes, its influence on unscheduled DNA synthesis, and its removal from the DNA of both cell types. We found that 20–30% of the BBA is bound to macromolecules other than DNA and that its binding to DNA is essentially complete after 30 min. The induction of unscheduled DNA synthesis by the carcinogen in XP lymphocytes was approximately 10% of that induced in normal lymphocytes. While 15–20% of the BBA was removed from the DNA of normal cells 6 h after treatment, only 1–2% was removed from the DNA of XP cells. Thus, XP cells not only are defective in repairing ultraviolet-damaged DNA and excising thymine dimers but also fail to repair DNA damaged by certain carcinogens, and, most importantly, fail to remove the DNA-bound carcinogen, BBA.     

Preparation of desiccation-resistant aquatic-living Nostoc flagelliforme (Cyanophyceae) for potential ecological application

Nostoc flagelliforme is a terrestrial edible cyanobacterium that grows in arid and semi-arid steppes. The continued over-exploitation in the last century has led to a sharp decline of this resource and a severe deterioration of the steppe ecology. Liquid-cultured N. flagelliforme serves as promising algal ‘seeds’ for resource restoration. In this study, macroscopic (or visible) aquatic-living colonies (MaACs) of N. flagelliforme were developed under weak light and high nitrogen conditions. In a 24 day shake-flask culture, MaACs were propagated by about 4.5-fold in biomass without loss of their macro-morphology; at the same time, the addition of weak UV-B treatment resulted in slightly bigger MaACs. Polyvinylpyrrolidone (PVP) k30, a water-soluble polymer, was used to generate the coating around MaACs, and after full desiccation, the coated MaACs could recover their photosynthetic physiological activity when rehydrated, with 4% PVP k30 for coating being most effective. In contrast, PVP k30-coated microscopic aquatic-living colonies of N. flagelliforme and non-coated MaACs showed no resistance to full desiccation. The macroscopic morphology or structure of MaACs should be crucial for the formation of protection by PVP k30 coating. PVP k30-coated MaACs were more approaching to actual application for resource restoration.