The selectivity of sexual responses to song displays: effects of partial chemical lesion of the HVC in female canaries

Cervicovaginal secretions were collected from 26 women (13 premenopausal and 13 postmenopausal) using a new sampling device (MucoSafeTM) with an absorbent which was introduced into the vagina and retrieved by the women themselves, after which it was air-dried and stored for months at room temperature until extraction of immunoglobulins. Cervical secretions were also collected by absorbent cylindrical wicks (Polyfiltronics) which were introduced into the cervical canal during speculum examination and thereafter kept frozen until extraction. The concentrations of specific IgA and IgG antibodies (to group B streptococci) in extracts from both methods were corrected by reference to total immunoglobulin levels. Three pairs of samples, all from postmenopausal women, were excluded from analysis due to undetectable levels of antibodies in the MucoSafeTM specimen. In the remaining 23 pairs, corrected concentrations of IgA and IgG antibodies in samples obtained by MucoSafeTM correlated well with the corresponding concentrations in wick samples, R = 0.84 (p < 0.0001) and R = 0.69 (p = 0.0002), respectively. Thus, cervicovaginal secretions for antibody measurements can be obtained by this novel method for self-sampling, obviating the need for speculum examination and storage of frozen samples.     

Effect of sample storage on quantitation of lipoprotein(a) by an enzyme-linked immunosorbent assay

This study evaluated the effect of storage on the quantitation of lipoprotein (Lp)(a) in 25 serum samples. Aliquots of serum were stored for up to three years at either -20 degrees C or -70 degrees C and Lp(a) subsequently analyzed using an enzyme-linked immunosorbent assay kit. Concentrations of Lp(a) declined during storage, and the temperatures employed elicited significantly different (P < 0.05) values within 12 mon which further diverged during three years of storage. Compared to baseline values, significant decreases (P < 0.05) in Lp(a) levels were evident after six months of storage at -20 degrees C with apparent losses (geometric mean) reaching 36.9% (95% confidence interval: 30.9%, 42.9%) after three years. Similarly, significantly lower (P < 0.05) Lp(a) values were recorded after six months of storage at -70 degrees C and at three years the decrease (geometric mean) was 19.1% (95% confidence interval: 14.3%, 24.0%). The losses, after three years, in terms of the arithmetic mean were 53.5 and 26.2% at -20 and -70 degrees C, respectively. Phenotype analysis suggested that large isoforms are more susceptible to degradation than smaller moieties. This may be related to the observation that apparent losses are reduced in samples containing over 8 mg/dL Lp(a). Nevertheless, Lp(a) levels in stored samples retained a strong correlation with the baseline values. These results must be considered specific for the storage conditions and analytical procedures employed.     

Antigenicity of a synthetic peptide from glucosyltransferases of Streptococcus mutans in humans

Human salivary immunoglobulin A (IgA) and serum IgG antibodies to the Streptococcus mutans glucosyltransferases (Gtfs) and to a synthetic peptide of 19 amino acids from a conserved region in the Gtfs (residues 435 to 453) were determined in young adults by enzyme-linked immunosorbent assay. Varying levels of antibody to Gtfs were detected in saliva or serum, with significantly higher levels of antibody to GtfD than to GtfB/C or GtfC. Anti-Gtf IgA levels in saliva did not correlate with those of IgG in serum. Caries-free (CF) volunteers exhibited significantly higher salivary IgA antibody levels to the peptide and to GtfB/C or GtfC than did the caries-active (CA) subjects. Preincubation of CF saliva and serum with the peptide inhibited the antibodies to the Gtfs in a dose-dependent manner, whereas preincubation of the samples from the CA group resulted in only partial inhibition. Our results indicated that this 19-amino-acid peptide includes one of the major B-cell epitopes of Gtfs and that CF individuals have higher titers of antibodies than CA subjects.     

Comparison of the oral, rectal, and vaginal immunization routes for induction of antibodies in rectal and genital tract secretions of women

To determine which mucosal immunization routes may be optimal for induction of antibodies in the rectum and female genital tract, groups of women were immunized a total of three times either orally, rectally, or vaginally with a cholera vaccine containing killed Vibrio cholerae cells and the recombinant cholera toxin B (CTB) subunit. Systemic and mucosal antibody responses were assessed at 2-week intervals by quantitation of CTB-specific antibodies in serum and in secretions collected directly from mucosal surfaces of the oral cavity, rectum, cervix, and vagina with absorbent wicks. The three immunization routes increased levels of specific immunoglobulin G (IgG) in serum and specific IgA in saliva to similar extents. Rectal immunization was superior to other routes for inducing high levels of specific IgA and IgG in rectal secretions but was least effective for generating antibodies in female genital tract secretions. Only vaginal immunization significantly increased both specific IgA and specific IgG in both the cervix and the vagina. In addition, local production of CTB-specific IgG in the genital tract could be demonstrated only in vaginally immunized women. Vaginal immunization did not generate antibodies in the rectum, however. Thus, generation of optimal immune responses to sexually transmitted organisms in both the rectal and the genital mucosae of women may require local immunization at both of these sites.     

Systemic and mucosal IgA responses to systemic antigen challenge in IgA nephropathy

IgA nephropathy (IgAN) is characterized by the deposition of glomerular IgA. The source of the deposited IgA is not known, but both the mucosal and systemic IgA systems have been implicated. In order to investigate mucosal and systemic antibody production to systemic antigen challenge in IgAN, 20 patients and 20 controls where immunized with tetanus toxoid (TT). While patients with IgAN responded with a similar serum IgG, IgA, IgA1, and IgA2 antibody response to controls, they did, however, produce more IgA1 antibodies relative to IgA2 (P < 0.05). No salivary IgA antibody response was observed to systemic immunization in controls; however, there was a significant IgA response to TT in the saliva of patients with IgAN. IgA antibodies were produced in vitro by Epstein Barr virus (EBV)-transformed peripheral blood lymphocytes (PBLs) obtained from control blood only when taken shortly (1 or 2 weeks) after immunization. Patients with IgAN produced significantly more IgA anti-TT positive cultures than controls and for a longer period (P < 0.01) after immunization. In contrast, IgG anti-TT was produced in EBV-transformed cultures at all time points, but with no difference between IgAN and controls in the proportion of IgG producing cultures. These results demonstrate increased IgA antibody production in both the systemic and mucosal IgA systems following systemic immunization in IgAN and suggest an abnormal overlap between the two systems in IgAN.     

Evidence for differential effects of sulphasalazine on systemic and mucosal immunity in rheumatoid arthritis

OBJECTIVE: To study the effects of sulphasalazine (SASP) on the systemic and mucosal humoral immune systems in patients with rheumatoid arthritis (RA). METHODS: Serum concentrations of interleukin 6 (IL-6), class and subclass specific IgG, IgA and IgM, IgA and IgG antigliadin antibodies and rheumatoid factors (RF) of IgG, IgA (including IgA1 and IgA2 subclasses) and IgM isotypes were measured before and 16 weeks after sulphasalazine (SASP) therapy in 15 female and three male patients with RA. Amounts of immunoglobulins in saliva and jejunal fluid were measured as estimates of mucosal humoral immunity. RESULTS: Serum concentrations of IgA and IgG decreased significantly during SASP therapy and correlated with reduced concentrations of IL-6. In addition, levels of circulating IgA RF, IgA anti-gliadin antibodies and IgM RF decreased significantly after the treatment. In contrast, immunoglobulin levels in saliva and jejunal fluid were unaltered. CONCLUSION: SASP exerts powerful but selective inhibitory effects on systemic immunoglobulin production, whereas no effects on mucosal immunoglobulin production were observed. The decreased systemic B cell activity may be mediated by downregulation of the production of IL-6, a cytokine with Ig switching properties.     

Mucosal antibody expression following rapid SIV(Mne) dissemination in intrarectally infected Macaca nemestrina

The early kinetics of antibody expression following transmucosal infection by SIV(Mne) were examined in several mucosal compartments in Macaca nemestrina. Five male-female pairs of macaques were inoculated intrarectally with SIV(Mne) E11S, a biological clone, and serially euthanized at 1, 2, 4, 8, and 12 weeks postinoculation. Plasma, tears, saliva, rectal secretions, and vaginal washes were collected serially and just prior to euthanasia. Both total and SIV-specific IgG and IgA levels were measured by immunoglobulin isotype-specific quantitative enzyme-linked immunosorbent assays (ELISAs), and were further examined by conventional and enhanced chemiluminescence (ECL) immunoblots. Virus coculture, polymerase chain reaction, and in situ hybridization assays revealed the systemic spread of virus as early as 1 week postinoculation in 8 of 10 animals. ECL immunoblots detected SIV-specific antibodies in mucosal samples collected 1 week postinoculation. The most dramatic increases in both total and SIV-specific IgA levels were detected in rectal secretion samples. In contrast, plasma and nonrectal mucosal samples from the same time points increased only slightly, suggesting that the most robust antibody response occurred at the portal of infection. Our results show that the SIV-infected macaque is an excellent model for studies designed to assess mucosal immune responses to primate lentivirus infections. Additional studies will assess the correlation between the antiviral protection afforded by candidate vaccines and mucosal antibody responses.     

Effect of different storage temperatures, sample collection procedures and immunoassay methods on osteocalcin measurement

The apparent instability of measured osteocalcin has been reported as method-dependent and related to preanalytical variables such as storage temperature, and the use of anticoagulants and protease inhibitors. The aim of this study was to determine a sample collection procedure which minimised osteocalcin degradation. Blood samples from five normal individuals were collected with or without anticoagulants and protease inhibitors (heparin, EDTA, or heparin and aprotinin) and stored at 4 degrees C, -20 degrees C or -70 degrees C for up to 7 days, 28 days and 90 days respectively. Osteocalcin was measured by both a monoclonal EIA specific for intact osteocalcin and a bovine polyclonal RIA. Osteocalcin concentrations in serum and EDTA-treated samples significantly decreased by 40% (P < 0.001) with the ELISA and 72% (P < 0.001) with the RIA after 7 days storage at 4 degrees C. Similar falls were documented in these samples when stored at -20 degrees C and -70 degrees C and measured by the ELISA. Minimal changes in osteocalcin immunoreactivity were observed in either assay when heparin-treated plasma with or without aprotinin was stored at -20 degrees C or -70 degrees C for up to 90 days. The apparent instability of measured osteocalcin can be minimised using these conditions.     

Role of sarcoplasmic reticulum in the contractile dysfunction during myocardial ischaemia and reperfusion

The aim of our study was to analyze HIV-specific humoral immunity in the intestinal mucosa at different stages of HIV infection in comparison with serum and saliva. Duodenal biopsy specimens from 30 AIDS patients and 9 HIV-infected patients without AIDS were cultured for 48 hours. Culture supernatants, as well as simultaneously obtained serum and saliva samples, were adjusted to the same immunoglobulin concentrations and tested for HIV-specific IgG and IgA by Western blot. The HIV antigen pattern differed clearly between IgA and IgG but was similar for each isotype independent of its origin (i.e., serum, saliva, or biopsy specimen supernatants). Short-term cultured duodenal biopsy specimens from HIV-infected patients at all stages produced predominantly IgG, which was broadly reactive with HIV antigens. Lower titers of HIV-specific IgA, which recognized few antigens, were found, mostly the glycoprotein gp160. At later stages of the disease compared with earlier stages, the reaction pattern of mucosal IgA from saliva and biopsy supernatants was even more restricted; secretory component was frequently absent. The abnormal predominance of HIV-specific IgG over IgA in mucosal secretions may result from abnormal antibody production in the mucosa rather than from serum leakage. Mucosal inflammation induced by HIV-IgG immune complexes and insufficient immune exclusion by secretory IgA may not only lead to increased mucosal HIV replication but may also contribute to gastrointestinal disease in HIV-infected patients.     

gamma/delta T cell-deficient mice have impaired mucosal immunoglobulin A responses

Mucosal tissues of mice are enriched in T cells that express the gamma/delta T cell receptor. Since the function of these cells remains unclear, we have compared mucosal immune responses in gamma/delta T cell receptor-deficient (TCRdelta-/-) mice versus control mice of the same genetic background. The frequency of intestinal immunoglobulin (Ig) A plasma cells as well as IgA levels in serum, bile, saliva, and fecal samples were markedly reduced in TCRdelta-/- mice. The TCRdelta-/- mice produced much lower levels of IgA antibodies when immunized orally with a vaccine of tetanus toxoid plus cholera toxin as adjuvant. Conversely, the antigen-specific IgM and IgG antibody responses were comparable to orally immunized control mice. Direct assessment of the cells forming antibodies against the tetanus toxoid and cholera toxin antigens indicated that significantly lower numbers of IgA antibody-producing cells were present in the intestinal lamina propria and Peyer's patches of TCRdelta-/- mice compared with the orally immunized control mice. The selective reduction of IgA responses to ingested antigens in the absence of gamma/delta T cells suggests a specialized role for gamma/delta cells in mucosal immunity.     

Energetics of vinca alkaloid interactions with tubulin isotypes: implications for drug efficacy and toxicity

Capture ELISAs, for canine IgG, IgM, IgA and albumin, were developed and used to analyse immunoglobulin (Ig) concentrations in both serum and secretions. Matched samples of serum, saliva and tears were taken from 31 dogs, assigned to two groups based on age, whilst bile samples were obtained from nine adult dogs at post-mortem. Serum and tear IgA concentrations were significantly lower in dogs < or = 12 months of age compared with dogs > 12 months of age (p = 0.006 and 0.045, respectively). There was no significant correlation between serum and secretory Ig levels, with the single exception of serum and tear IgM concentrations (rp = 0.553, p = 0.004). IgG and IgM concentrations were significantly correlated in matched tear and saliva samples (IgG: rp = 0.470, p = 0.023; IgM: rp = 0.651, p < 0.0001). Albumin concentrations were significantly correlated with IgG, but not IgM or IgA, in both saliva and tears (saliva, rp = 0.581, p = 0.004; tears, rp = 0.843, p < 0.0001) whilst IgA and IgM concentrations were significantly correlated with each other in both secretions (saliva, rp = 0.644, p = 0.001; tears, rp = 0.555, p = 0.009). Significantly, more Ig of all classes was secreted into saliva than tears as calculated by a secretory index. A large diurnal and day-to-day variation was observed in Ig concentrations in serial saliva and tear samples taken from a further four dogs. Serum Ig concentrations are therefore, poor indicators of mucosal secretion in this species and significant intra-individual variation exists in secretory Ig levels. Both findings should be taken into account in future studies of canine mucosal immunoglobulins.     

Disease surveillance in the ED: factors leading to the underreporting of gonorrhea

Viable periodontal ligament (PL) cells are required for PL healing of avulsed teeth following replantation. If immediate replantation cannot be accomplished, the ability of PL progenitor cells to reproduce (clonogenic capacity) and recolonize the wound may be extended by prevention of desiccation and storage in physiological media. This investigation examined the effects of storage in saliva, milk, Hank's balanced salt solution (HBSS) and Eagle's medium (alpha MEM) on the clonogenic capacity of human PL progenitor cells at 30 and 60 min extra-alveolar time. Twenty erupted human premolar teeth extracted as atraumatically as possible for orthodontic purposes were used in the present study. Fifteen premolars were placed immediately in freshly collected autologous saliva at room temperature, (+ 23 degrees C) for 15 min. These 15 premolars were next divided into three groups of five and stored in either saliva, milk or HBSS at + 4 degrees C in plastic cups surrounded by ice. The remaining five teeth served as positive controls and were immediately placed in alpha MEM at + 4 degrees C. PL tissue was scraped from one-half of the root surface with a scalpel at 30 and 60 min total extra-alveolar duration. Cells were released from the tissue sample with a 30 min enzymatic digestion procedure and the cells from the tissue samples analyzed for clonogenic capacity. There was a reduction in clonogenic capacity with time for all protocols. Periodontal ligament cells stored in alpha MEM showed the least reduction between 30 and 60 min and the greatest reduction was observed for PL cells stored in saliva. The difference in clonogenic capacity following transfer from saliva to milk or HBSS was not significant at 30 min. At 60 min, cells transferred from saliva to HBSS had a statistically higher percentage of clonogenic cells than those transferred to milk (5.9% vs. 3.5%; P < 0.05). We conclude that immediate storage of avulsed teeth in autologous saliva, followed by transfer to chilled milk, preserves the presence of sufficient progenitor cells in the PL to warrant replantation and the possibility of PL healing at 60 min extra-alveolar duration.     

Analytical methods and stability assessment of liquid yeast derived sucrase

Two independent analytical methods for determining the activity and stability profile of liquid yeast derived sucrase (YS) were established and validated in order to conduct preliminary stability studies as a function of temperature. The methods included a hexokinase-based (HK) enzymatic assay for determining the formation of glucose upon hydrolysis of sucrose by YS, and a direct polarimetric procedure to quantitate YS hydrolysis of sucrose. Both assays were validated with respect to YS dilution, incubation time, sucrose or glucose concentration, linearity of response and within- and between-day variability. A preliminary stability study was conducted over a 24 week period with liquid YS samples stored at -20, 4, 30, 40 and 50 degrees C. Enzymatic activity was monitored as a function of time using both the HK and polarimetric assays. Liquid YS samples stored at -20, 4 and 30 degrees C retained 100% activity after 24 weeks storage, while the samples stored at 40 degrees C lost approximately 70% activity over the same storage period and samples stored at 50 degrees C lost approximately 95% activity after 12 weeks storage. The two methods of analysis gave consistent results over the course of the study.     

Storage temperature of the unbuffered rapid urease test

OBJECTIVES: The unbuffered rapid urease test (RUT) is an accurate, rapid, and inexpensive method for detecting Helicobacter pylori. However, it is generally recommended that the reagent be prepared daily. This prospective study was undertaken to evaluate the shelf life of our unbuffered RUT when stored at 4 and -20 degrees C. METHODS: Ninety-five patients were studied. Three sets of antral (X2) and body (X1) biopsy samples were taken from each patient. The samples were subjected to histological examination, with the RUTs stored at 4 and -20 degrees C. The RUT tubes were examined at 1 and 15 min. RESULTS: Fifty-six patients (59%) were infected with H. pylori as defined by histological examination. The reagent was classified according to storage time (group I, < or = 5 days; group II, > 5 days). The mean (SD) storage time of group I (n = 59) and group II (n = 36) was 3.2 (1.4) and 9.9 (5.0) days, respectively. At 15 min, the sensitivity of our RUT stored at 4 degrees C was significantly higher in group I than in group II (92 vs 47%). On the other hand, the sensitivity of our RUT stored at -20 degrees C remained consistently high in both groups (15 min: group I, 92%; group II, 100%). Our RUTs stored at 4 and -20 degrees C were highly specific in both groups. CONCLUSIONS: Our RUT remains highly sensitive and specific when it is stored at 4 degrees C for up to 5 days. When the RUT is expected to be stored for a longer period of time, the bottles should be frozen at -20 degrees C.     

Role of mucosal IgA in the resistance to Acanthamoeba keratitis

PURPOSE: To determine whether oral immunization with Acanthamoeba castellanii antigens elicits mucosal antibodies of the IgA isotype and whether mucosal antibodies affect parasite adhesion to the corneal epithelium. METHODS: Chinese hamsters were immunized with 100 microg aqueous Acanthamoeba antigen mixed with cholera toxin (Ac-CT) and subsequently challenged with parasite-laden contact lenses that were applied to abraded corneal surfaces. Tears and stool samples were examined for the presence of Acanthamoeba-specific IgA antibodies by enzyme-linked immunosorbent assay (ELISA). The effect of mucosal antibody on trophozoite binding to corneal epithelium and viability of trophozoites was examined in vitro. RESULTS: Hamsters immunized orally with Ac-CT showed significantly lower infection rates than did control groups (21.4% versus 72.6%). ELISA analysis of mucosal specimens showed the presence of parasite-specific IgA in stool samples and tears from hamsters orally immunized with Ac-CT, but not in control animals. In vitro assays showed that anti-Acanthamoeba IgA did not affect parasite viability. However, mucosal anti-Acanthamoeba IgA profoundly inhibited (>75%) the binding of parasites to corneal epithelial cells in vitro. CONCLUSIONS: Oral immunization with Ac-CT induces the production of parasite-specific IgA in mucosal secretions and prevents corneal infection. Mucosal antibody does not affect the viability of Acanthamoeba trophozoites but seems to prevent infection by inhibiting parasite binding to the corneal epithelium.     

Solitary scapula mass: atypical presentation of esophageal adenocarcinoma

The stability of adenosine in various diluents in polypropylene syringes and polyvinyl chloride (PVC) bags at three temperatures was studied. Portions of pooled undiluted adenosine infusion (3 mg/ mL) were stored in 60-mL capped syringes, 20 for each storage condition. Adenosine infusions were prepared by mixing adenosine with 5% dextrose injection, 0.9% sodium chloride injection, lactated Ringer's injection, or 5% dextrose and lactated Ringer's injection to produce a concentration of 0.75 mg/mL. Samples of each infusion were stored in 60-mL capped syringes and 50-mL bags, 20 syringes and 20 bags for each storage condition. Syringes and bags were stored in the dark at 25, 5, and -15 degrees C. At various sampling times, three syringes and three bags of each infusion were removed for visual inspection, pH measurement, and high-performance liquid chromatographic analysis. At 10 and 16 days, fungal growth at 25 degrees C was suspected in the infusions prepared with 5% dextrose injection. For all other samples, there was no evidence of precipitation or change in pH. The concentration of adenosine remained constant in all samples at all storage conditions. Adenosine 3 mg/mL was stable in polypropylene syringes for 7 days at 25 degrees C, 14 days at 5 degrees C, and 28 days at -15 degrees C; adenosine 0.75 mg/ mL in 0.9% sodium chloride injection and in 5% dextrose injection was stable in polypropylene syringes and PVC bags for 16 days at 25, 5, and -15 degrees C; and adenosine 0.75 mg/mL in lactated Ringer's injection and in 5% dextrose and lactated Ringer's injection was stable in syringes and bags for 14 days at 25, 5, and -15 degrees C.     

Novel intranasal immunization techniques for antibody induction and protection of mice against gastric Helicobacter felis infection

Intranasal (i.n.) delivery of antigen can be highly effective for generating circulating and secretory antibody responses. Mice were immunized i.n. with two antigens, human IgA, and Helicobacter pylori urease in the presence or absence of mucosal adjuvant. To restrict antigen delivery to the upper airways, protein solutions were administered in a small volume without anesthesia. Repeated daily i.n. administration of antigen without adjuvant elicited high levels of specific IgG in serum and IgA in serum, saliva, and feces. Once weekly i.n. immunization with co-administration of cholera toxin or Escherichia coli heat-labile toxin as adjuvant elicited somewhat lower levels of antibody to urease. When challenged with Helicobacter felis, only mice immunized with urease in the presence of adjuvant were protected against gastric infection.     

Regeneration across preserved peripheral nerve grafts

The potential to store nerve grafts for a prolonged period of time was assessed in a rat sciatic nerve model. Three-centimeter syngeneic nerve grafts were stored in Belzer/University of Wisconsin cold storage solution at different temperatures (5 degrees C, 22 degrees C, or 37 degrees C) for varying time periods (6 h, 24 h, or 3 weeks) prior to transplantation. Functional assessment using serial walking track analyses revealed no difference between storage times and temperatures. At 14 months postengraftment, the conduction velocities and the number of myelinated fibers that had regenerated across all grafts stored at 5 degrees C for all time periods tested were superior to grafts stored at either 22 degrees C or 37 degrees C. Nerve grafts stored for up to 3 weeks at 5 degrees C acted as effective conduits for proximal regenerating fibers and resulted in histologic, electrophysiologic, and functional results equivalent to fresh nerve grafts. Nerve graft storage may be applicable to nerve allografts and potentially provide allograft material that requires reduced or no associated host immunosuppression.     

Effects of vocal loudness on nasalance measures

The effect of storage and high temperatures on the stability of Saccharomyces cerevisiae allergens was studied by immunoblotting. Saccharomyces cerevisiae allergic serum pool and 125I- and galactosidase-labelled anti-IgE were used in the assays. Freeze-dried extracts were reconstituted with saline and with 50% glycerol and then stored at room (+20 degrees C) and refrigerator temperature (+6 degrees C) for different time periods. The stability was better in 50% glycerol at +6 degrees C than at room temperature without glycerol. However, after 1 month, two of the most important allergens of S. cerevisiae, the 48 and 32 kDa protein allergens, lost their IgE-binding capacity even in the extracts stored with 50% glycerol at +6 degrees C. The 45 kDa allergen was, on the other hand, quite stable after storage for 9 months at +6 degrees C. Although the beneficial effect of 50% glycerol was clear, storage at +6 degrees C, even with 50% glycerol should not exceed 1 month for S. cerevisiae extracts. Two commercially available S. cerevisiae extracts in solution with valid expiry dates were also analysed. They had only little allergenic potency, while a freeze-dried extract stored for 8 years showed good allergenic potency. Heating S. cerevisiae extracts resulted in precipitation, the precipitated fraction contained almost all the specific proteins as judged by electrophoresis and IgE detection. The supernatant fraction contained only a few allergens.     

In vitro production of cyanide in normal human blood and the influence of thiocyanate and storage temperature

Normal human blood stored at room temperature (about 20 degrees C) may, over a period of weeks, undergo a slow transformation of its cyanide content. In contrast, when normal blood is stored at -20 degrees C there is formation of cyanide, usually occurring most rapidly during the first few days of storage. Peak concentrations are never greater than 20 microgram/100 ml, and although some fluctuation in concentration occurs over several months of storage at deep freeze temperature, levels are always higher than in the freshly drawn blood. The amount of cyanide produced appears to be a function of both the initial thiocyanate concentration and the freezing and/or thawing of blood. Blood stored at refrigerator temperature (about 4 degrees C) has the least fluctuation in cyanide concentration over a storage period up to three months. During this study it was found that there is significant difference in whole blood cyanide concentration between smokers and nonsmokers.