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Inpatient hospital admission and death after outpatient surgery in elderly patients: importance of patient and system characteristics and location of care.

HYPOTHESIS: Surgery at different outpatient care locations in the higher-risk elderly (age >65 years) population is associated with similar rates of inpatient hospital admission and death. DESIGN: Claims analysis of patients undergoing 16 different surgical procedures in a nationally representative (5%) sample of Medicare beneficiaries for the years 1994 through 1999. SETTING: Hospital-based outpatient centers, freestanding ambulatory surgery centers (ASCs), and physicians' office facilities. PATIENTS: Medicare beneficiaries older than 65 years. MAIN OUTCOME MEASURES: Rate of death, emergency department risk, and admission to an inpatient hospital within 7 days of outpatient surgery. RESULTS: We studied 564,267 outpatient surgical procedures: 360,780 at an outpatient hospital, 175,288 at an ASC, and 28,199 at a physician's office. There were no deaths the day of surgery at a physician's office, 4 deaths the day of surgery at an ASC (2.3 per 100,000 outpatient procedures), and 9 deaths the day of surgery at an outpatient hospital (2.5 per 100,000 outpatient procedures). The 7-day mortality rate was 35 per 100,000 outpatient procedures at a physician's office, 25 per 100,000 outpatient procedures at an ASC, and 50 per 100,000 outpatient procedures at an outpatient hospital. The rate of admission to an inpatient hospital within 7 days of outpatient surgery was 9.08 per 1000 outpatient procedures at a physician's office, 8.41 per 1000 outpatient procedures at an ASC, and 21 per 1000 outpatient procedures at an outpatient hospital. In multivariate models, more advanced age, prior inpatient hospital admission within 6 months, surgical performance at a physician's office or outpatient hospital, and invasiveness of surgery identified those patients who were at increased risk of inpatient hospital admission or death within 7 days of surgery at an outpatient facility. CONCLUSION: This study represents an initial effort to demonstrate the risk associated with outpatient surgery in a large, diverse population of elderly individuals.

Age Factors↗

2001 Consensus Guidelines for the management of women with cervical cytological abnormalities.

OBJECTIVE: To provide evidence-based consensus guidelines for the management of women with cervical cytological abnormalities and cervical cancer precursors. PARTICIPANTS: A panel of 121 experts in the diagnosis and management of cervical cancer precursors, including representatives from 29 professional organizations, federal agencies, and national and international health organizations, were invited to participate in a consensus conference sponsored by the American Society for Colposcopy and Cervical Pathology (ASCCP). EVIDENCE AND CONSENSUS PROCESS: Guidelines for the management of women with cervical cytological abnormalities were developed through a multistep process. Starting 6 months before the conference, working groups developed draft management guidelines based on formal literature reviews of English-language articles published in 1988-2001, as well as input from the professional community at large, obtained using interactive Internet-based bulletin boards. On September 6-8, 2001, the ASCCP Consensus Conference was held in Bethesda, Md. Guidelines with supporting evidence were presented and underwent discussion, revision, and voting. CONCLUSIONS: Management of women with atypical squamous cells (ASC) depends on whether the Papanicolaou test is subcategorized as of undetermined significance (ASC-US) or as cannot exclude high-grade squamous intraepithelial lesion (HSIL) (ASC-H). Women with ASC-US should be managed using a program of 2 repeat cytology tests, immediate colposcopy, or DNA testing for high-risk types of human papillomavirus (HPV). Testing for HPV DNA is the preferred approach when liquid-based cytology is used for screening. In most instances, women with ASC-H, low-grade squamous intraepithelial lesion, HSIL, and atypical glandular cells should be referred for immediate colposcopic evaluation.

Adolescent↗

Alteration of nitric oxide synthase activity upon oxidative stress in cultured retinal cells.

In the present study we examined the effect of oxidative stress-mediated hydroperoxide formation on the activity of nitric oxide synthase (NOS) in retinal cells in culture. Oxidative stress was induced in the presence of Fe2+ and ascorbate or Fe2+ alone and compared to H2O2-induced maximal cellular oxidation, and was measured by following the formation of intracellular hydroperoxides with the probe DCFH2 (2',7'-dichlorodihydrofluorescein). After a 15-min exposure to the oxidants, formation of hydroperoxides was significantly increased in the presence of 100 microM Fe2+ (about twofold), as compared to the control. Coadministration of Fe2+ and ascorbate (Fe-Asc) did not affect DCF fluorescence, but highly reduced the intracellular pH (pHi = 6.32 +/- 0.08), in comparison with control conditions (pHi = 7.05 +/- 0.11), as determined with the probe BCECF (2',7'-bis-(carboxyethyl)-5(and-6) carboxyfluorescein). Nevertheless, preincubation of Fe-Asc at acidic pH also increased the formation of hydroperoxides. Oxidative stress induced in the presence of Fe-Asc (at pH 6.5) significantly decreased the activity of NOS by 20% of control activity, as determined by the formation of [14C]citrulline. Fe-Asc (pH 6.5) also reduced the production of cyclic GMP (cGMP) in retinal cells by 1.5-fold, although a decrement in pH from 7.4 to 6.5 was not sufficient to decrease cGMP production. These data suggest that NO. production may be compromised in the presence of Fe-Asc. Moreover, neither 4 mM dithiotreitol (DTT) nor 4 mM glutathione (GSH) altered the production of cGMP in retinal cells submitted to oxidative stress. A reduction in NO. generation upon oxidative stress may reduce major damaging effects induced by ONOO- in cultured retinal cells.

Animals↗

Inflamed kidneys of NZB / W mice are a major site for the homeostasis of plasma cells.

(NZB x NZW)F1 (NZB / W) mice develop a disease similar to human systemic lupus erythematosus (SLE), including autoantibody production, hypergammaglobulinaemia and inflammation of the kidneys. It is known that large numbers of lymphocytes infiltrate the kidneys of these mice. Here, we compare the roles of bone marrow, spleen and inflamed kidneys of NZB / W mice in the activation of B cells and the persistence of antibody-secreting cells (ASC). ASC are present in the kidneys of NZB / W mice with full-blown disease, as many as in the spleen and bone marrow. The specificity of the ASC in the inflamed kidneys is not restricted to self-antigens. After immunization of NZB / W mice with ovalbumin (OVA) the OVA-specific ASC are found initially in the spleen. Weeks later, OVA-specific ASC are found in high numbers in the bone marrow and the kidneys of these mice, but no longer in the spleen. As determined by FACS, B cells with a germinal center phenotype (B220(+) / PNA(+)) are found only in very low numbers in the kidneys, but in high numbers in the spleen of NZB / W mice. Germinal centers could not be detected in the kidneys, but in the spleen, and plasma cells appear to be scattered over the tissue. These data suggest that in autoimmune NZB / W mice, plasma cells generated in immune reactions of secondary lymphoid organs, later accumulate and persist in the inflamed kidneys, were they enhance the local concentrations of Ab and immunocomplexes. These experiments identify the inflamed kidneys of NZB / W mice as a site of prime relevance for the homeostasis of plasma cells, irrespective of their specificity.

Animals↗

NH2-SEP-PAK cartridges for enrichment of Aromatic Sulfur Compounds from sea water: determination by GC-FID and GC-MS.

The Aromatic Sulfur Compounds (ASCs) are considered as "markers" of the crude oil age. In this frame, we have considered ASCs as "markers" of remote crude oil sea water pollution for their chemical characteristics, good solubility in water and resistance to the biodegradation. Gas chromatography of ASCs in distilled water and sea water samples has been performed after adsorption from a 80 mL water sample on to a cartridge containing 100 mg NH2-bonded porous silica. The ASCs are desorbed with 2 mL acetone-trichloromethane (1:1), which is concentrated and analysed by GC-FID or GC-MS in SIM mode. The average recovery of 0.1 microg mL(-1) of each ASC from distilled water and 0.04-0.2 microg mL(-1) from sea water samples is > or = 96.3% with a standard deviation < or = 2.2. The limits of detection are 0.06-0.05 ng mL(-1) for thiophen-2-carboxaldehyde and benzothiophene, and 0.006-0.004 ng mL(-1) for dibenzothiophene and thiantrene in GC-FID whereas they are 0.02 ng mL(-1) for thiophen-2-carboxaldehyde and benzothiophene, and 0.003 ng mL(-1) for dibenzothiophene and thiantrene in GC-MS (SIM) with a relative standard deviation < or =7.

Adsorption↗

Dehydroascorbate transport in human chondrocytes is regulated by hypoxia and is a physiologically relevant source of ascorbic acid in the joint.

OBJECTIVE: To evaluate the dehydroascorbate (DHA) transport mechanisms in human chondrocytes. METHODS: The transport of L-(14)C-DHA in human chondrocytes was analyzed under various conditions, including the use of RNA interference (RNAi), to determine the role of glucose transporter 1 (GLUT-1) and GLUT-3 in L-14C-DHA transport and to evaluate the effects of physiologically relevant oxygen tensions on L-14C-DHA transport. In order to estimate the contributions of reduced ascorbic acid (AA) and DHA to intracellular ascorbic acid (Asc), the quantities of AA and DHA were measured in synovial fluid samples from osteoarthritis (OA) patients and compared with the reported levels in rheumatoid arthritis (RA) patients. RESULTS: DHA transport in human chondrocytes was glucose-sensitive, temperature-dependent, cytochalasin B-inhibitable, modestly stereoselective for L-DHA, and up-regulated by low oxygen tension. Based on the RNAi results, GLUT-1 mediated, at least in part, the uptake of DHA, whereas GLUT-3 had a minimal effect on DHA transport. DHA constituted a mean 8% of the total Asc in the synovial fluid of OA joints, in contrast to 80% of the reported total Asc in RA joints. CONCLUSION: We provide the first evidence that chondrocytes transport DHA via the GLUTs and that this transport mechanism is modestly selective for L-DHA. In the setting of up-regulated DHA transport at low oxygen tensions, DHA would contribute 26% of the total intracellular Asc in OA chondrocytes and 94% of that in RA chondrocytes. These results demonstrate that DHA is a physiologically relevant source of Asc for chondrocytes, particularly in the setting of an inflammatory arthritis, such as RA.

Arthritis, Rheumatoid↗

Atypical squamous cells of undetermined significance--rule out high-grade squamous intraepithelial lesion: cytopathologic characteristics and clinical correlates.

The significance and clinical management of atypical squamous cells of undetermined significance (ASCUS) on cervical cytologic smears has been an area of much controversy. This study compiled a list of criteria useful in identifying the subset of cases that would be categorized as atypical squamous cells of undetermined significance- rule out high-grade squamous intraepithelial lesion (ASC-H) in the new Bethesda System terminology, which eventuate in a diagnosis of cervical intraepithelial neoplasia (CIN). The cytopathology files at Johns Hopkins Hospital were searched for ASC-H cases from the 3-yr period 1996-1998, which had definitive clinicopathologic follow-up (colposcopy and cervical biopsies). The smears were reviewed, cytomorphologic features studied, and clinical correlations performed. ASC-H was diagnosed in 257 of 45,428 gynecologic smears (0.6%), 72 having had clinicopathologic follow-up. Of these 72 cases, 35 (49%) on follow-up had a negative/reactive diagnosis (NR), whereas 37 (51%) turned out to be CIN [CIN-I-18 (49%) and CIN II and III-19 (51%)]. The significant cytomorphologic differences in the ASC-H category with a CIN follow-up (compared with an NR follow-up) were fewer atypical cells, more often discohesive or seen singly, more monomorphic, a higher nuclear-to-cytoplasmic (N/C) ratio, greater nuclear hyperchromasia, more coarse, unevenly dispersed chromatin, more prominent nuclear membrane irregularities, lack of nucleoli, chromocenters or nuclear grooves, and lack of an inflammatory background. Careful attention to subtle cytomorphologic characteristics may be helpful for a more definitive subdivision of ASC-H terminology into a NR and a CIN diagnosis.

Female↗

p16(INK4A) is a surrogate biomarker for a subset of human papilloma virus-associated dysplasias of the uterine cervix as determined on the Pap smear.

Recently, p16(INK4A) has been identified as a biomarker for human papilloma virus (HPV)-induced dysplastic lesions of the cervix and it has been suggested that it may be a useful diagnostic aid for these lesions. This study therefore was performed to determine the utility of p16 expression in a series of Papanicolaou (Pap) smears collected in liquid medium and to determine its benefit, if any, over HPV testing. One hundred seven cases, including 23 negative cases, 34 with low-grade squamous intraepithelial lesion (LSIL), 16 with high-grade squamous intraepithelial lesion (HSIL), 29 with atypical squamous cells of uncertain significance (ASC-US), and 5 cases with ASC suspicious for HSIL (ASC-H), were evaluated for both p16 expression and HPV DNA. We observed p16 expression in only 36% of all cases with abnormal cytology (30/84) and in 40% of all cases associated with high-risk HPV. The highest rate of positivity (80%) and the highest levels of expression (more than three to five positive cells/10x field) were seen in HSIL. Similar results were observed with ASC-H cases. This suggests that in equivocal cases, p16 may be used for confirmation of the diagnosis. On the other hand, p16 positivity was noted in only 21% of LSIL and ASC-US cases. This raises the interesting possibility, given that only a minority of LSIL cases progress on to higher-grade lesions, that p16 might be useful for triaging these patients for closer follow-up and/or further evaluation. Additional studies are required for confirmation.

Biomarkers, Tumor↗

Differential expression of cdk inhibitors p16, p21cip1, p27kip1, and cyclin E in cervical cytological smears prepared by the ThinPrep method.

Our objective was to correlate p16, p21cip1, p27kip1, and cyclin E protein expression with the degree of dysplasia on ThinPrep Papanicolaou (Pap) smears using a modified immunoperoxidase staining. Smears read as normal, atypical squamous cells of undetermined significance (ASC-US), low-grade squamous intraepithelial lesion (LSIL), or high-grade SIL (HSIL) were identified and tested for high-risk human papillomavirus (HR-HPV). Additional smears were processed for immunoperoxidase for p16, p21cip1, p27kip1, and cyclin E. Thirty-four smears were satisfactory for study. The p16 was positive in all nine HSIL, in four of nine LSIL, and in one of seven ASC-US. The p27kip1 was positive in all nine HSIL, in eight of nine LSIL, and in one of seven ASC-US. The p21cip1 was positive in all nine HSIL, in one of nine LSIL, and in one of seven ASC-US. Cyclin E was positive in seven of nine HSIL and in one of nine LSIL and in none of the ASC-US smears. Normal smears were negative for all the antigens. There was poor correlation of protein expression and HR-HPV infection. We concluded that p16, p21cip1, p27kip1, and cyclin E can be demonstrated on Pap smears and they are expressed differentially in dysplastic cells, with highest expression in HSIL. The p21cip1 and cyclin E showed the greatest correlation with HSIL.

Cell Cycle Proteins↗

Atypical squamous cells, cannot exclude a high-grade squamous intraepithelial lesion: the practice experience of a hospital-based reference laboratory with this new Bethesda system diagnostic category.

The 2001 Bethesda System diagnostic category of atypical squamous cells cannot exclude a high grade squamous intraepithelial lesion (ASC-H) was developed and implemented after many studies that proved its clinical relevance. In this study, we describe the practice experience of a hospital-based reference laboratory with this new diagnostic category. The anatomic pathology computer database was searched, and 414 Papanicolaou (Pap) smears signed out as ASC-H were discovered among 60,390 Pap smear accessions over a 17-month period of time. One hundred four cases had corresponding tissue specimens. Slides from all Pap smears and tissue specimens were reviewed, and five Pap smears were reclassified, leaving 99 study cases. In our laboratory, 88.9% of the study cases had either low or high grade dysplasia diagnosed on subsequent tissue specimens. The positive predictive value of ASC-H for histologically proven high-grade squamous intraepithelial lesions (HSIL's) was 40.4%. Of those having human papillomavirus (HPV) hybrid capture II testing, high-risk HPV types were detected in 73.9% of cases. The majority of study cases had less than 25 atypical cells. In two hysterectomy cases and three loop electrosurgical excession procedure (LEEP) conization cases, high-grade dysplasia was present as a single microscopic focus, suggesting that the paucity of atypical cells in ASC-H Pap smears may be secondary to small lesion sampling. Thirteen study patients were postmenopausal and 30.8% had low-grade dysplasia, and of these, 46.2% had high-grade dysplasia on subsequent tissue specimens. In conclusion, our practice experience with ASC-H is similar to that reported in the literature before the 2001 Bethesda System.

DNA Probes, HPV↗

Impact of reflex HPV testing on interpretation and management of ThinPrep Pap tests.

HPV testing for high-risk types is the preferred method to triage women with atypical squamous cells (ASC) on ThinPrep Pap tests for management. The impact of this practice on the cytopathologists' interpretation is unknown. A significant percentage of providers at the Beth Israel Deaconess Medical Center (BIDMC) are not requesting reflex HPV testing. This offers a unique opportunity to examine its impact. Consecutive ThinPreps submitted to the cytology laboratory at the BIDMC for a 3-mo period were identified retrospectively. The ThinPreps were divided into those with a request for reflex HPV testing (reflex testing request group), those without such a request, and those with a request for HPV testing regardless of the diagnosis. Cytologic diagnoses, results of HPV testing and subsequent cervical biopsies within 6 mo after the ThinPrep were recorded and correlated in these cases. A total of 4,590 ThinPreps were processed during this period. 2,659 had a request for reflex HPV testing, 1,849 did not have such a request, and 82 had a request for HPV testing regardless of diagnosis. The numbers (%) of abnormal results in these three groups were 155 (5.8%), 141 (7.6%), and 22 (27%), respectively. The ASC:SIL ratios for these three groups were 1.2, 0.86, and 0.62, respectively. 38/71 (54%) women with ASC in the reflex testing request group were positive for high-risk HPV types. Of those with ASC in the reflex testing request and no request groups, 15/73 (21%) and 12/56 (21%) underwent biopsies with 2 (of 15) and 2 (of 12) showing HSIL on histology, respectively. Our study demonstrates that at our institution a request for reflex HPV testing did not significantly affect cytopathologists' interpretation of ThinPreps. Furthermore, HPV testing did not seem to have influenced the subsequent management of women with ASC diagnosis.

Biopsy↗

The use of antibody-secreting cell probes to reveal tissue-restricted immune responses during infection.

Antibody secreting cell probes (ASC-probes) were generated from the hepatic lymph node (HLN), mesenteric lymph node (MLN) and spleen of rats after infection with the liver fluke, Fasciola hepatica, and used to probe Western blots of parasite antigens. In chronic primary infections, parasite-specific antibodies were only detected in ASC-probes derived from HLN. Seven days after a secondary infection, a restricted response was detected in ASC-probes from the MLN, directed predominantly against an antigen specific to the newly excysted juvenile (NEJ) stage. This NEJ-specific antigen was only recognized by HLN if the second infection was not rejected and the challenge flukes reached the liver. Measurement of the immunoglobulin levels present in the ASC-probes showed significant elevation only in lymph nodes draining sites of recent infection. In addition, when the isotype profiles were determined in ASC-probes derived from different lymph nodes, it was observed that they showed different isotype preferences, in particular IgA for the MLN, IgE for HL.N and IgM for spleen. These results show that discrete and independent immune responses occur in different body compartments of a rat against different stages of a parasite.

Animals↗

Estimation of the potential for nitrosation and its inhibition in subjects from high- and low-risk areas for esophageal cancer in southern China.

We studied 2 counties in southern China, which are close to each other but show very different mortality rates from esophageal cancer. We collected 12 hr urine samples and analyzed them for NPRO, other nitrosamino acids, nitrate and ASC. The potential for forming NPRO and the effect on inhibition by ASC were recorded. Urine was collected after (i) no treatment, (ii) a dose of proline and (iii) proline with ASC. Intake and excretion of nitrate were significantly greater in the high-risk area. After the proline dose, urinary excretion of NPRO was significantly greater in the high-risk area. The inhibitory effect of ASC after taking it with proline was less in the high-risk area. Hence, the high-risk area showed greater potential for forming NPRO, and this formation was less inhibited by ASC. This supports the view that esophageal cancer in this area may be caused by nitrosamines formed in vivo, perhaps from nitrite produced by reduction of ingested nitrate.

China↗

Quantitative determination of water-soluble scavengers in neoplastic and non-neoplastic human breast tissue.

The water-soluble scavengers ascorbic acid (Asc), cysteine (Cys), glutathione (GSH) and uric acid (UA) as well as DNA content were determined in 40 breast tissue samples (neoplastic and non-neoplastic tissues from 20 patients). To allow proper homogenization to take place, a fixed number of sections was cut from a tissue cylinder of known diameter. Adjacent sections were stained with hematoxylin-eosin and the fractions of epithelium, fat and connective tissue were estimated as a percentage of the section area. Protein-free extracts were injected into a reverse-phase high-pressure liquid chromatography system and scavengers quantified with 2 electrochemical detectors (gold and glassy carbon). DNA and all scavengers, except UA, were greatly increased in cancer tissues in nearly all cases. Amounts of Asc and GSH in neoplastic tissue correlated closely with DNA values and percentage of epithelium, those of Cys not so closely and those of UA not at all. We assume that Asc and GSH were located mainly in the epithelium, UA mainly in the extracellular space and Cys in both spaces. When values were expressed as mumol/g DNA, a parameter related to content per cell, values were higher in neoplastic than in non-neoplastic tissue for Asc (18/20 cases), GSH (17/20) and Cys (14/20) and lower in neoplastic tissue for UA (19/20). It is known that increased GSH protects cells against certain drugs in tissue cultures. For in vivo treatment the presence of increased Asc (and to a lesser extent Cys) in addition to GSH could be of importance.

Adult↗

Neutral amino acid transport in embryonal carcinoma cells.

Neutral amino acid transport was characterized in the pluripotent embryonal carcinoma (EC) cell line, OC15. Ten of the thirteen amino acids tested are transported by all three of the major neutral amino acid transport systems--A, L, and ASC--although one system may make a barely measurable contribution in some cases. The characterization of N-methyl-aminoisobutyric acid (meAIB) transport points to this model amino acid as a definitive substrate for System A transport by OC15 cells. Thus, high concentrations of meAIB can be used selectively to block System A transport, and the transport characteristics of meAIB represent system A transport. Kinetic analysis of System A, with a Km = 0.79mM and Vmax = 14.4 nmol/mg protein/5 min, suggests a single-component transport system, which is sensitive to pH changes. While proline transport in most mammalian cells is largely accomplished through System A, it is about equally divided between Systems A and ASC in OC15 cells, and System A does not contribute at all to proline transport by F9 cells, an EC cell line with limited developmental potential. Kinetic analysis of System L transport, represented by Na+-independent leucine transport, reveals a high-affinity, single-component system. This transport system is relatively insensitive to pH changes and has a Km = 0.0031 mM and Vmax = 0.213 nmol/mg protein/min. The putative System L substrate, 2-aminobicyclo-[2,2,1]heptane-2-carboxylic acid (BCH), inhibits Systems A and ASC as well as System L in OC15 cells. Therefore, BCH cannot be used as a definitive substrate for System L in OC15 cells. Phenylalanine is primarily transported by Na+-dependent Systems A and ASC (83% Na+-dependent; 73% System ASC) in OC15 cells, while it is transported primarily by the Na+-independent System L in most other cell types, including early cleavage stage mouse embryos and F9 cells. We have also found this unusually strong Na+-dependency of phenylalanine transport in mouse uterine blastocysts (82% Na+-dependent). There is no evidence for System N transport by OC15 cells, since histidine is transported primarily by a Na+-independent, BCH-inhibitable mechanism.

Amino Acids↗

L-ascorbic acid 2-phosphate stimulates collagen accumulation, cell proliferation, and formation of a three-dimensional tissuelike substance by skin fibroblasts.

Proliferation of human skin fibroblasts was stimulated significantly by the presence of L-ascorbic acid 2-phosphate (Asc 2-P). The presence of Asc 2-P (0.1-1.0 mM) in the culture medium for 3 weeks enhanced the relative rate of collagen synthesis to total protein synthesis 2-fold as well as cell growth 4-fold. Coexistence of L-azetidine 2-carboxylic acid (AzC), an inhibitor of collagen synthesis, attenuated both effects of Asc 2-P in a dose-dependent manner. Supplementation of the medium with Asc 2-P also accelerated procollagen processing to collagen and deposition of collagen in the cell layer. Among the acidic glycosaminoglycans (GAG), another major component of extracellular matrix (ECM), deposition of sulfated forms was increased by the additive. Electron microscopic observations showed multilayered, rough endoplasmic reticulum-rich cells surrounded by dense ECM. These results indicate that Asc 2-P is useful in culture systems as a long-acting vitamin C derivative and also that it promotes reorganization of a three-dimensional tissuelike substance from skin fibroblasts in culture by stimulating collagen accumulation in the fibroblasts.

Ascorbic Acid↗

Light-induced ascorbate-dependent electron transport and membrane energization in chloroplasts of bundle sheath cells of the C4 plant maize.

Chloroplasts in bundle sheath cells (BSC) of maize perform photosystem I (PSI)-mediated production of ATP. In this study, the participation of ascorbate (Asc) as an electron donor to PSI in light-induced electron transport in isolated maize BSC was demonstrated. It was found that Asc, at physiological concentrations, rapidly reduced photooxidized reaction center chlorophyll of PSI (P700). The rate of Asc donation of electrons to P700+ reached rates of 50-100 microequivalents (mg Chl)(-1) h(-1) at 70-80 mM ascorbate with methyl viologen as an electron acceptor. Electron transport supported by Asc was coupled with membrane energization, as demonstrated by the light-induced formation of a trans-thylakoid electric field measured by the electrochromic shift of carotenoids. The possible physiological function of Asc-dependent electron transport in bundle sheath chloroplasts of maize, as an electron donor for linear electron flow versus sustaining cyclic electron transport, is discussed.

Ascorbic Acid↗

The gill is a major organ for antibody secreting cell production following direct immersion of sea bass (Dicentrarchus labrax, L.) in a Photobacterium damselae ssp. piscicida bacterin: an ontogenetic study.

Extremely high numbers of antibody secreting cells (ASC) were observed in the gills of sea bass fry immunised at three different age/sizes (initial weight of 0.1, 2 and 5 g) by direct immersion in a Photobacterium damselae spp. piscicida bacterin. The relatively low ASC production in the head kidney and spleen suggests that the systemic compartment was only slightly stimulated upon immersion vaccination. There was no response of corresponding magnitude in the gut as the one observed in the gills. A clear age effect was observed in the ASC response of the different groups, especially visible in the gills. Significantly higher numbers of specific ASC were observed in the gills of the two oldest groups (initial weight of 2 and 5 g) compared with the youngest fish (initial weight of 0.1 g), but the oldest groups were not significantly different from each other. Additionally, a more rapid response was observed with the ageing of the fish, with peak responses in all the organs at day 18, 16 and 8 post-immunisation in the smallest to largest fish, respectively. There was no evidence that direct immersion exposure to P. damselae ssp. piscicida at the earliest stages used in the present study (0.1 g) was tolerogenic. In the context of present knowledge, this study strongly supports the importance of the route of immunisation to locally stimulate ASC and the importance that the gills might have in specific responses.

Age Factors↗