A comparison of in vitro flowers to in vivo flowers of haploid and diploidNicotiana tabacum L.

Thin cell layers (TCLs) were cultured from inflorescences of diploid (2n=4x=48) and haploid (2n=2x=24)Nicotiana tabacum L. "Samsun" and the subsequent flowers formed in vitro were then compared to in vivo flowers. Plants derived from TCLs possessed flowers that were typical of their seed or androgenetically-derived counterparts, whereas de novo flowers from TCLs were abnormal when compared to their counterparts. The TCLs of haploid plants produced more flower buds than diploid TCLs, and did so in a shorter period of time. In vitro flowers and anthers at both ploidy levels were considerably smaller than the in vivo flowers; in vitro flowers also had variable numbers of anthers and pistils. The embryogenic capacity of anthers taken from in vivo diploid flowers was 5 times greater than that of in vitro diploid or haploid anthers. In vivo haploid anthers produced no embryoids, whereas in vitro haploid anthers did produce embryoids. Observations of mitotic cells in root tips of plants derived from anther cultures of in vitro haploid flowers revealed a mixoploid nature. Diploid meiosis was regular and haploid meiosis was irregular regardless of the origin (in vitro or in vivo) of the flowers.Supported by state Hatch funds.     

Monoclonal antibodies for use with 125iodine-labeled radioligands in progesterone radioimmunoassay

The interactions of heterologous and homologous 125I-iodinated radioligand with polyclonal and monoclonal antibodies directed against 11-hydroxyprogesterone hemisuccinate conjugated to bovine serum albumin were compared. Our data show that, with a polyclonal antibody, the use of the same bridge in the tracer and the antigen results in a low sensitivity while a heterologous tracer can decrease markedly the titer of antibody but increases the sensitivity of the radioimmunoassay. Due to the high specificity of monoclonal antibodies, the interaction with heterologous and homologous iodinated tracers is extremely variable from one antibody to another but the present results demonstrate that radioimmunoassays of very high sensitivity can be obtained with homologous and heterogeneous tracers. However, an appropriate tracer has to be selected to meet the requirement of such an assay.     

Factors affecting variability in anther culture and in conversion of androgenic embryos ofSolanum phureja

The variation for embryo production in anther ofSolanum phureja was examined as a function of maximum greenhouse temperature prior to bud harvest and innate responsiveness among anthers within a bud. Four clones ofS. phuyreja were grown in a greenhouse under a 16-h photoperiod. The temperature was monitored continuously. Buds (60 per day on 10 days) were collected and the anthers cultured in two groups of five flasks (30 anthers per flask). In the first group, each flask contained the 30 anthers from six buds; in the second group, each flask contained one anther from each of 30 buds. Significantly smaller coefficients of variation were observed for the second group, suggesting that variation for embryogenic capcity among buds was greater than that among anthers within a bud. Variation in embryo yield as a function of greenhouse temperature was examined by stepwise regression analysis. Embryogenic capacity of one clone was adversely affected by high temperatures (31–37°C) that occurred two and seven days before bud harvest. However, similarly high temperatures appeared to enhance the androgenic response of another clone. Conversion of anther-derived embryos over three subcultures to fresh regeneration medium was examined as a function of anther donor or clone, cold pretreatment of embryos, and morphological classification of embryos. Only clonal origin significantly affected conversion rate which ranged from 12.5% to 46.0%. Conversion rate declined on each serial subculture.Abbreviations BA N⁶-benzyladenine - GA₃ gibberellic acid, IAA-indole-3-acetic acid     

Stimulation of DNA repair by the spermatidal TP1 protein

The chromatin remodeling process that takes place during spermiogenesis in mammals is characterized by a transient increase in DNA single-strand breaks (SSB). The mammalian transition proteins (TPs) are expressed at a high level at mid-spermiogenesis steps coincident with chromatin remodeling and could be involved in the repair of these lesions since SSB are no longer detected in terminally differentiated spermatids. We report that TP1 can stimulate the repair of SSB in vitro and demonstrate that in vivo repair of UV-induced DNA lesions is enhanced in mammalian cells stably expressing TP1. These results suggest that, aside from its role in DNA compaction, this major transition protein may contribute to the yet unidentified enzymatic activity responsible for the repair of SSB at mid-spermiogenesis steps. These results also suggest that the TP1 proteins have the potential to participate in the repair process following genotoxic insults and therefore may play an active role in the maintenance of the integrity of the male haploid genome during spermiogenesis.     

Plasma kinetics of VLDL and HDL apoC-I in normolipidemic and hypertriglyceridemic subjects

ApoC-I has several different lipid-regulating functions including, inhibition of receptor-mediated uptake of plasma triglyceride-rich lipoproteins, inhibition of cholesteryl ester transfer activity, and mediation of tissue fatty acid uptake. Since little is known about the rate of production and catabolism of plasma apoC-I in humans, the present study was undertaken to determine the plasma kinetics of VLDL and HDL apoC-I using a primed constant (12 h) intravenous infusion of deuterium-labeled leucine. Data were obtained for 14 subjects: normolipidemics (NL, n = 4), hypertriglyceridemics (HTG, n = 4) and combined hyperlipidemics (CHL, n = 6). Plasma VLDL triglyceride (TG) levels were 0.59 +/- 0.03, 4.32 +/- 0.77 (P < 0.01 vs. NL), and 2.20 +/- 0.39 mmol/l (P < 0.01 vs. NL), and plasma LDL cholesterol (LDL-C) levels were 2.34 +/- 0.22, 2.48 +/- 0.26, and 5.35 +/- 0.48 mmol/l (P < 0.01 vs. NL), respectively. HTG and CHL had significantly (P < 0.05) increased levels of total plasma apoC-I (12.5 +/- 1.2 and 12.4 +/- 1.3 mg/dl, respectively) versus NL (7.9 +/- 0.6 mg/dl), due to significantly (P < 0.01) elevated levels of VLDL apoC-I (5.8 +/- 0.8 and 4.5 +/- 0.8 vs. 0.3 +/- 0.1 mg/dl). HTG and CHL also had increased rates of VLDL apoC-I transport (i.e., production) versus NL: 2.29 +/- 0.34 and 3.04 +/- 0.53 versus 0.24 +/- 0.11 mg/kg.day (P < 0.01), with no significant change in VLDL apoC-I residence times (RT): 1.16 +/- 0.12 versus 0.69 +/- 0.06 versus 0.74 +/- 0.17. Although HDL apoC-I concentrations were not significantly lower in HTG and CHL versus NL, HDL apoC-I rates of transport were inversely related to plasma and VLDL-TG levels (r = -0.63 and -0.62, respectively, P < 0.05). Our results demonstrate that increased levels of plasma and VLDL apoC-I in hypertriglyceridemic subjects (with or without elevated LDL-C levels) are associated with increased levels of plasma VLDL apoC-I production.     

CLUSTER ANALYSIS OF ANTHER-DERIVED PLANTS OF SOLANUM PHUREJA (SOLANACEAE) BASED ON MORPHOLOGICAL CHARACTERISTICS

The feasibility of grouping anther-derived plants of Solarium phureja according to ploidy based on their morphological characteristics was studied. Canonical discriminant analysis identified four characteristics (anther length, number of chloroplasts per pair of guard cells, leaf width, corolla width at widest diam) of nine measured as the most effective combination for diagnosing ploidy. Data for these characteristics from two sets of plants were subjected to two clustering techniques, one using the average linkage clustering (UPGMA of the NT-SYS programs) and the other using centroid sorting (SAS-Fastclus). Screening of anther-derived plants by cluster analysis proved to be an efficient means of separating monoploids from the other ploidy levels.     

Visual acuity in the cathemeral strepsirrhine Eulemur macaco flavifrons

Studies of visual acuity in primates have shown that diurnal haplorhines have higher acuity (30–75 cycles per degree (c/deg)) than most other mammals. However, relatively little is known about visual acuity in non‐haplorhine primates, and published estimates are only available for four strepsirrhine genera (Microcebus, Otolemur, Galago, and Lemur). We present here the first measurements of visual acuity in a cathemeral strepsirrhine species, the blue‐eyed black lemur (Eulemur macaco flavifrons). Acuity in two subjects, a 3‐year‐old male and a 16‐year‐old female, was assessed behaviorally using a two‐alternative forced choice discrimination task. Visual stimuli consisted of high contrast square wave gratings of seven spatial frequencies. Acuity threshold was determined using a 70% correct response criterion. Results indicate a maximum visual acuity of 5.1 c/deg for the female (1718 trials) and 3.8 c/deg for the male (846 trials). These values for E. macaco are slightly lower than those reported for diurnal Lemur catta, and are generally comparable to those reported for nocturnal Microcebus murinus and Otolemur crassicaudatus. To examine ecological sources of variation in primate visual acuity, we also calculated maximum theoretical acuity for Cheirogaleus medius (2.8 c/deg) and Tarsius syrichta (8.9 c/deg) using published data on retinal ganglion cell density and eye morphology. These data suggest that visual acuity in primates may be influenced by activity pattern, diet, and phylogenetic history. In particular, the relatively high acuity of T. syrichta and Galago senegalensis suggests that visual predation may be an important selective factor favoring high visual acuity in primates. Am. J. Primatol. 71:343–352, 2009. © 2009 Wiley‐Liss, Inc.