Denitrification and total nitrate uptake in streams of a tropical landscape.

Rapid increases in nitrogen (N) loading are occurring in many tropical watersheds, but the fate of N in tropical streams is not well documented. Rates of nitrate uptake and denitrification were measured in nine tropical low-order streams with contrasting land use as part of the Lotic Intersite Nitrogen eXperiment II (LINX II) in Puerto Rico using short term (24-hour) additions of K(15)NO3 and NaBr. Background nitrate concentrations ranged from 105 to 997 microg N/L, and stream nitrate uptake lengths were long, varying from 315 to 8480 m (median of 1200 m). Other indices of nitrate uptake (mass transfer coefficient, V(f) [cm/s], and whole-stream nitrate uptake rate, U [microg N m(-2) s(-1)]) were low in comparison to other regions and were related to chemical, biological, and physical parameters. Denitrification rates were highly variable (0-133 microg N m(-2) min(-1); median = 15 microg N m(-2) min(-1)), were dominated by the end product N2 (rather than N2O), and were best predicted by whole-stream respiration rates and stream NO3 concentration. Denitrification accounted for 1-97% of nitrate uptake with five of nine streams having 35% or more of nitrate uptake via denitrification, showing that denitrification is a substantial sink for nitrate in tropical streams. Whole-stream nitrate uptake and denitrification in our study streams closely followed first-order uptake kinetics, indicating that NO3 uptake is limited by delivery of substrate (NO3) to the organisms involved in uptake or denitrification. In the context of whole-catchment nitrogen budgets, our finding that in-stream denitrification results in lower proportional production of N2O than terrestrial denitrification suggests that small streams can be viewed as the preferred site of denitrification in a watershed in order to minimize greenhouse gas N2O emissions. Conservation of small streams is thus critical in tropical ecosystem management.

Evaluation of the ARMD1 locus on 1q25-31 in patients with age-related maculopathy: genetic variation in laminin genes and in exon 104 of HEMICENTIN-1.

The age-related maculopathy (ARM) genetics program at Columbia University utilizes comprehensive genetic analysis of candidate genes in large case-control studies to determine genotypes associated with the ARM complex trait. Genes encoding laminins, a class of extracellular matrix proteins, represent attractive candidates for two reasons. First, the presence of laminins in the basal lamina of the retinal pigment epithelium (RPE), Bruch's membrane, and choriocapillaris suggests a possible role in the pathophysiology of ARM. Second, three laminin genes, LAMC1, LAMC2, and LAMB3, are located in the 1q25-31 region, within the previously mapped ARMD1 locus. The entire open reading frame of the three laminin genes was screened for variants by denaturing high-performance liquid chromatography (DHPLC) and direct sequencing in at least 92, and up to 368 ARM patients and matched unaffected controls. Sixty-nine sequence variants were detected in the 69 exons of the LAMC1, LAMC2, and LAMB3 genes. Screening of exon 104 of the recently proposed ARMD1 gene, HEMICENTIN-1, residing in the 1q25-31 locus, did not detect the suggested causal variant, Q5345R, in 632 study subjects. Overall, we did not find statistically significant differences in the frequency of variants between ARM-affected individuals and age-matched controls. Four rare, non-synonymous, variants were detected in single cases of ARM patients. Our data on relatively limited numbers of study subjects do not suggest a significant role for genetic variation in the three laminin genes and in exon 104 of HEMICENTIN-1 in predisposing individuals to ARM. However, as in many instances in similar studies, involvement of rare amino acid-changing variants in a fraction of ARM cannot be ruled out.

Change on the horizontal and vertical meridians of the cornea after cataract surgery.

PURPOSE: To compare the course and magnitude of change on the horizontal and vertical meridians of the cornea after 5 different incisions for cataract: extracapsular cataract extraction (ECCE), 6 mm superior scleral tunnel (6Sup), 3 mm superior scleral tunnel (3Sup), 3 mm temporal scleral tunnel (3Temp), and 3 mm temporal corneal incision (3Cor). METHODS: Retrospective chart review of 665 cases of preoperative regular astigmatism. The preoperative keratometry (K) reading was subtracted from the postoperative K reading to determine mean net change on each meridian at 1 day, 1 week, 2 weeks, 1 month, 1.5 months, 2 months, 4 months, 6 months and 12 months and at 6 month intervals thereafter. After the superior incisions, the temporal changes on each meridian are well described by an analytic model with an initial and final plateau. The changes after the temporal incisions are described by a linear equation. RESULTS: After each superior incision, the steepness and length of the transition from the initial to final plateau for each meridian depend on incision length. Considering the uncertainty of measuring K, the corneal meridians stabilized 4.5 months after ECCE, 1.2 months after 6Sup, and 0.3 months after 3Sup. No significant change was detected on the horizontal and vertical meridians after 3Temp and 3Cor. CONCLUSION: The magnitude and the duration of changes on the horizontal and vertical meridians of the cornea after cataract surgery depend on both incision length and location. Small temporal incisions induce less change than superior incisions.

An action spectrum for UV-B radiation and the rat lens.

PURPOSE: To determine an action spectrum for UV-B radiation and the rat lens and to show the effect of the atmosphere and the cornea on the action spectrum. METHODS: One eye of young female rats was exposed to 5-nm bandwidths of UV-B radiation (290, 295, 300, 305, 310, and 315 nm). Light scattering of exposed and nonexposed lenses was measured 1 week after irradiation. A quadratic polynomial was fit to the dose-response curve for each wave band. The dose at each wave band that produced a level of light scattering greater than 95% of the nonexposed lenses was defined as the maximum acceptable dose (MAD). Transmittance of the rat cornea was measured with a fiberoptic spectrophotometer. The times to be exposed to the MAD in Stockholm (59.3 degrees N) and La Palma (28 degrees N) were compared. RESULTS: Significant light scattering was detected after UV-B at 295, 300, 305, 310, and 315 nm. The lens was most sensitive to UV-B at 300 nm. Correcting for corneal transmittance showed that the rat lens is at least as sensitive to UV radiation at 295 nm as at 300 nm. The times to be exposed to the MAD at each wave band were greater in Stockholm than in La Palma, and in both locations the theoretical time to be exposed to the MAD was least at 305 nm. CONCLUSIONS: After correcting for corneal transmittance, the biological sensitivity of the rat lens to UV-B is at least as great at 295 nm as at 300 nm. After correcting for transmittance by the atmosphere, UV-B at 305 nm is the most likely wave band to injure the rat lens in both Stockholm and La Palma.

Age-related changes in the absorption characteristics of the primate lens.

PURPOSE: To quantitate aging of the primate lens by changes in the absorption characteristics that are related to the yellowing of lens protein. METHODS: The lenses of lower primates and humans were sectioned anterior to posterior every 0.25 mm, and the UV-visible spectrum of each section was measured to determine the cumulative spectra along the visual axis. The ratio of the absorbance at 320 nm (formed with aging) to the absorbance at 365 nm (present in the young lens) was correlated with the age of the lens. RESULTS: In the young primate UV-B is transmitted to the retina, and UV-A is transmitted to the nucleus of the lens. By puberty, changes in the absorption characteristics of the lens that are associated with the yellowing of lens protein prevented most of the UV-B from reaching the retina and by the eighth decade, the transmittances at 320 and 365 nm to the nucleus of the lens were approximately 40% and 79%, respectively. A linear relationship between the ratio of absorbance at 320 to 365 nm and age was found for both lower primates and humans to the age of 80 years. This is surprising, because the maximum life span of the lower primate is approximately 35 years, whereas humans may live 100 years. CONCLUSIONS: These data suggest that the observed spectral changes associated with the yellowing of the lens are the result of a chronological process, such as chemical or photochemical modifications, not biological aging.

Transmission spectra of light to the mammalian retina.

A simple method has been developed to determine the optical properties of the anterior segment of the intact eye. This consists of a probe that is inserted into the posterior sclera and detects light passing through the anterior segment. The probe is connected to a charge-coupled device spectrophotometer via a fiber optic bundle. It was determined that the young rat anterior segment transmits light down to 300 nm, whereas calf and rabbit eyes transmit no UVB and only part of the UVA to the posterior segment. The absorbing species in these animals is most likely NAD(P)H, which has an absorption maximum at approximately 345 nm and is associated with zeta-crystallin. A young primate anterior segment transmits almost no UV with a steep increase in transmission at CA 400 nm. Because some experiments employed a light tube that is used to illuminate the retina during vitrectomies, this method can be used to determine the transmission spectra of the anterior segment of humans in vivo.

Managing an imaging department with a business perspective. A view from the other side.

I came to the position of director of radiology at Winchester Hospital with two advanced degrees, in business administration and in health-care administration, to replace a director of 35 years, who had come from a technical background. That this community hospital outside Boston nourishes the entrepreneurial spirit is one of the premises underlying its success. I was supported from the beginning both by the radiologists, who supported the concept of a business-person running the department, and by the manager, whose acceptance of my role helped set the tone for the department. My goal was to create a vision for the department, which I could do only by establishing a relationship with the supervisory staff. Communication became my greatest challenge: to listen and learn from my staff and to articulate how we could move to a new level of performance. My management philosophy included giving budget responsibility to the supervisors, to teach how revenue and expense directly impact the department's business. We learned together the importance of customer service and how it, too, could impact our business. Finally, I helped supervisors offer their own ideas to solve department problems. Joining the AHRA, reading its publications and attending its conferences have helped me gain a broader perspective of the industry. Now, I see my role as a facilitator who sets a vision for the department and provides the proper support, tools and guidance for the staff to achieve results. Together, we have created a successful balance between the business and the patient care components.

The optical properties of the anterior segment of the eye: implications for cortical cataract.

Epidemiological studies have correlated cortical cataract with exposure to light and have suggested that this is due primarily to relatively short wavelengths of ultraviolet radiation (UV-B). In addition, some cellular and animal models also implicate UV-B. In order to evaluate the likely role of different wavelengths of light in the etiology of cortical cataracts, the optical characteristics of several animal models were ascertained and compared to the primate. This study shows that the mouse model absorbs UV-B almost exclusively whereas other animal models such as the rabbit and the guinea pig also contain chromophores that absorb UV-A. The absorptive characteristics of the human lens varies drastically with age. The young lens absorbs primarily UV-A, whereas with age, there are increases in absorptions at 320 nm and out to wavelengths as long as 550 nm. By sectioning human lenses it was found that these changes in absorption properties increased toward the central and the nuclear regions. These absorptive characteristics were then compared to the amount of light reaching the surface of the lens. It was found that UV-B is a minor component of total energy reaching the surface of the human lens and old human lens proteins absorb 2 orders of magnitude more UV-A and visible light than UV-B. It is concluded that it is premature to exclude UV-A or even visible light in the etiology of human cortical cataracts.

The Drosophila activin receptor baboon signals through dSmad2 and controls cell proliferation but not patterning during larval development.

The TGF-beta superfamily of growth and differentiation factors, including TGF-beta, Activins and bone morphogenetic proteins (BMPs) play critical roles in regulating the development of many organisms. These factors signal through a heteromeric complex of type I and II serine/threonine kinase receptors that phosphorylate members of the Smad family of transcription factors, thereby promoting their nuclear localization. Although components of TGF-beta/Activin signaling pathways are well defined in vertebrates, no such pathway has been clearly defined in invertebrates. In this study we describe the role of Baboon (Babo), a type I Activin receptor previously called Atr-I, in Drosophila development and characterize aspects of the Babo intracellular signal-transduction pathway. Genetic analysis of babo loss-of-function mutants and ectopic activation studies indicate that Babo signaling plays a role in regulating cell proliferation. In mammalian cells, activated Babo specifically stimulates Smad2-dependent pathways to induce TGF-beta/Activin-responsive promoters but not BMP-responsive elements. Furthermore, we identify a new Drosophila Smad, termed dSmad2, that is most closely related to vertebrate Smads 2 and 3. Activated Babo associates with dSmad2 but not Mad, phosphorylates the carboxy-terminal SSXS motif and induces heteromeric complex formation with Medea, the Drosophila Smad4 homolog. Our results define a novel Drosophila Activin/TGF-beta pathway that is analogous to its vertebrate counterpart and show that this pathway functions to promote cellular growth with minimal effects on patterning.

The Nop60B gene of Drosophila encodes an essential nucleolar protein that functions in yeast.

The Cbf5 protein of Saccharomyces cerevisiae was originally identified as a low-affinity centromeric DNA-binding protein, and chf5 mutants have a defect in rRNA synthesis. A closely related protein from mammals, NAP57, is a nucleolar protein that coimmunoprecipitates with the nucleolar phosphoprotein Nopp140. To study the function of this protein family in a higher eukaryote that is amenable to genetic approaches, the gene encoding a Drosophila melanogaster homolog, Nop60B, was identified. The predicted Drosophila protein shares a high degree of sequence identity over a 380-residue region with both the mammalian and yeast proteins, and shares several conserved motifs with the prokaryotic tRNA pseudouridine 55 synthases. Nop60B RNA is found at high levels in nurse cells and in the oocyte, and is present throughout development. Nop60B protein is localized primarily to the nucleolus of interphase cells, and is absent from the chromosomes during mitosis. Nop60B mutants were generated and shown to be homozygous lethal. The Drosophila gene can rescue the lethal phenotype of yeast chf5 mutations, showing that the function of this protein has been conserved from yeast to Drosophila.

Ribosomal protein insufficiency and the minute syndrome in Drosophila: a dose-response relationship.

Minutes comprise > 50 phenotypically similar mutations scattered throughout the genome of Drosophila, many of which are identified as mutations in ribosomal protein (rp) genes. Common traits of the Minute phenotype are short and thin bristles, slow development, and recessive lethality. By mobilizing a P element inserted in the 5' UTR of M(3)95A, the gene encoding ribosomal protein S3 (RPS3), we have generated two homozygous viable heteroalleles that are partial revertants with respect to the Minute phenotype. Molecular characterization revealed both alleles to be imprecise excisions, leaving 40 and 110 bp, respectively, at the P-element insertion site. The weaker allele (40 bp insert) is associated with a approximately 15% decrease in RPS3 mRNA abundance and displays a moderate Minute phenotype. In the stronger allele (110 bp insert) RPS3 mRNA levels are reduced by approximately 60%, resulting in an extreme Minute phenotype that includes many morphological abnormalities as well as sterility in both males and females due to disruption of early gametogenesis. The results show that there is a correlation between reduced RPS3 mRNA levels and the severity of the Minute phenotype, in which faulty differentiation of somatic tissues and arrest of gametogenesis represent the extreme case. That heteroalleles in M(3)95A can mimic the phenotypic variations that exist between different Minute/rp-gene mutations strongly suggests that all phenotypes primarily are caused by reductions in maximum protein synthesis rates, but that the sensitivity for reduced levels of the individual rp-gene products is different.

Analysis of the Legionella pneumophila fliI gene: intracellular growth of a defined mutant defective for flagellum biosynthesis.

Using a PCR-based strategy and degenerate oligonucleotides, we isolated a Legionella pneumophila gene that showed high sequence similarity to members of the fliI gene family. An insertion mutation that disrupted the fliI open reading frame was recombined onto the L. pneumophila chromosome and analyzed for its effects on production of flagella and intracellular growth. The mutation resulted in loss of surface-localized flagellin protein but had no effect on the ability of the bacteria to grow within cultured cells. Therefore, in spite of the fact that some aflagellar mutations render L. pneumophila unable to grow within macrophages, the isolation of this defined mutant confirms that production of flagella is not required for intracellular growth.

Cytoplasmically anchored plakoglobin induces a WNT-like phenotype in Xenopus.

Plakoglobin is one of two vertebrate proteins closely related to the Drosophila segment polarity gene product armadillo. Overexpression of plakoglobin induces neural axis duplication in Xenopus and the exogenous plakoglobin is localized to nuclei (Karnovsky, A., and Klymkowsky, M. W., Proc. Natl. Acad. Sci. USA 92, 4255, 1995; Rubenstein, A., et al., Dev. Genet., 1997, in press). We have carried out a series of experiments to test whether the nuclear localization of plakoglobin is required for its inductive effects. Prior to the midblastula transition exogenous plakoglobin is cytoplasmic and concentrated in the cortical regions of blastomeres; after the midblastula transition exogenous plakoglobin accumulates in embryonic nuclei. The addition of a "nuclear localization sequence" does not change the timing of plakoglobin's nuclear localization, suggesting that it is anchored in the cytoplasm prior to the midblastula transition. Next, we constructed two "membrane-anchored" forms of plakoglobin. These are exclusively cytoplasmic; yet both were as effective at producing a "Wnt-like" axis duplication as were "free," unfettered forms of plakoglobin. Moreover, expression of anchored plakoglobins had no apparent effect on the cytoplasmic or nuclear levels of beta-catenin. These data indicate that plakoglobin can act cytoplasmically to generate a WNT-like phenotype. Taken together with the ventralizing effects of a mutant from of the XTcf-3 transcription factor, described by Molenaar et al. Cell 86, 391, 1996, we speculate that in the early Xenopus embryo, activation of plakoglobin (or beta-catenin) inhibits the activity of XTcf-3 or a XTcf-3-like factor.

Fine-needle aspiration diagnosis of enterogenous cyst of the orbit: a case report.

Enterogenous cyst is a benign lesion derived from misplaced endodermal epithelium. We report the aspiration cytology findings of an orbital cyst from a 32-year-old woman. The smears contain benign-appearing cuboidal glandular cells with focal mucinous features. Carcinoembryonic antigen level in the cyst fluid was markedly elevated. These findings are consistent with recurrence of the enterogenous cyst initially diagnosed 7 years earlier.

Interactions of Drosophila Ultrabithorax regulatory regions with native and foreign promoters.

The Ultrabithorax (Ubx) gene of the Drosophila bithorax complex is required to specify parasegments 5 and 6. Two P-element "enhancer traps" have been recovered within the locus that contain the bacterial lacZ gene under the control of the P-element promoter. The P insertion that is closer to the Ubx promoter expresses lacZ in a pattern similar to that of the normal Ubx gene, but also in parasegment 4 during embryonic development. Two deletions have been recovered that remove the normal Ubx promoter plus several kilobases on either side, but retain the lacZ reporter gene. The lacZ patterns from the deletion derivatives closely match the normal pattern of Ubx expression in late embryos and imaginal discs. The lacZ genes in the deletion derivatives are also negatively regulated by Ubx and activated in trans by Contrabithorax mutations, again like the normal Ubx gene. Thus, the deleted regions, including several kilobases around the Ubx promoter, are not required for long range interactions with Ubx regulatory regions. The deletion derivatives also stimulate transvection, a pairing-dependent interaction with the Ubx promoter on the homologous chromosome.

Localizing the adhesive and signaling functions of plakoglobin.

Plakoglobin (PKG) is a major component of cell-cell adhesive junctions. It is also closely related to the Drosophila segment polarity gene product armadillo and can induce a WNT-like neural axis duplication (NAD) phenotype in Xenopus [Kamovsky and Klymkowsky, 1995.] To define the regions of PKG involved in cell adhesion and inductive signaling, we examined the behavior of mutated forms of PKG in Xenopus. Deletion of amino acids 22 through 39 (in the Xenopus PKG sequence increased the apparent stability of the polypeptide within the embryo and increased its ability to induce a WNT-like, NAD phenotype when expressed in the vegetal hemisphere. The N-terminal "head" and first 6 "ARM" repeats of PKG, or the C-terminal "tail" and the last 3 "ARM" repeats, could be removed without destroying the remaining polypeptide's ability to induce a NAD phenotype. The nuclear localization of mutant PKGs, however, was not strictly correlated with the ability to induce a NAD phenotype, i.e., some inactive polypeptides still accumulate in nuclei. Removal of PKG's head and first ARM repeat, which includes its alpha-catenin binding site, resulted in a polypeptide that, when expressed in the embryo, generated alpha dramatic cell adhesion defect. Removal of the next three ARM repeats abolished this adhesion defect, suggesting that the polypeptide no longer competes effectively with endogenous catenins for binding to cadherins. Expression of a form of PKG truncated after the 5th ARM repeat produced a milder cell adhesion defect, whereas expression of a polypeptide truncated after the 8th ARM repeat had little apparent effect on cellular adhesion. Based on these observations, we conclude that functions related to stability and cellular adhesion reside in the N-terminal region of the polypeptide, whereas the ability to induce a NAD phenotype lies within repeats 6-10 of the central region. The function(s) of the C-terminal domain of PKG remain uncertain at this time.

Astigmatism and visual recovery after 'large incision' extracapsular cataract surgery and 'small' incisions for phakoemulsification.

PURPOSE: This study compares the change over time of the astigmatism caused by "large" incision extracapsular cataract extraction (ECCE) and three smaller incisions for phakoemulsification. Based on this data, a mathematical model that predicts the course of astigmatism after a superior incision of length 3 to 12 mm has been developed. The relationship of axial length and preoperative astigmatism to induced post-operative astigmatism, the recovery of visual acuity, and the rate of YAG laser capsulotomy after each procedure also are documented. METHODS: Induced astigmatic change was calculated using a simple method of vector analysis. The change in induced astigmatism was calculated for 8 years after ECCE (n = 144), for 3 years after 6 mm superior incisions (6SUP) (n = 93), for 2 years after 3 mm superior incisions (3SUP) (n = 120), and for 18 months after 3 mm temporal incisions (3Temp) (n = 65). Plotted semi-logarithmically, the astigmatic change in each group may be represented mathematically. RESULTS: Two weeks after ECCE the mean induced cylinder was +3.47 D, which decayed to about -1.25 D after 6 months. Induced cylinder increased gradually to about -1.6 D after 8 years, although this further change was not significantly different than that at 6 months after surgery. For the phako groups, the net induced cylinder on the first post-operative day was: +1.23 D (6SUP), +0.49 D (3Sup), and -0.19 D (3Temp). After 6Sup the wound was astigmatically stable after approximately 3 months, and 3 years after surgery net induced cylinder was -0.66 D. After 3Sup the wound was astigmatically stable after about 6 weeks, and after 18 months net induced cylinder was -0.35 D. No significant change in astigmatism was detected at any time after 3Temp. Maximum visual acuity was reached after a mean of approximately 6 weeks after ECCE, 2 weeks after 6Sup, and between 1 day and 1 week after 3Sup and 3Temp. The rate of YAG laser capsulotomy was higher after ECCE than after any of the phakoemulsification procedures. No relationship of axial length or preoperative astigmatism to astigmatic change was detected. CONCLUSIONS: Incision size and location affect post-operative astigmatism. Induced astigmatism decreases with wound size, and only the 3 mm temporal incision is astigmatically neutral. The time for visual recovery increases with wound size. There appears to be less need for laser capsulotomy after phakoemulsification with capsulorrhexis than after ECCE. Axial length does not affect induced astigmatism after any of the 4 incisions, and preoperative astigmatism does not affect astigmatic change after ECCE and 6Sup.