Nuclear singlet relaxation by scalar relaxation of the second kind in the slow-fluctuation regime.

The singlet state of nuclear spin-1/2 pairs is protected against many common relaxation mechanisms. Singlet order, which is defined as the population difference between the nuclear singlet and triplet states, usually decays more slowly than the nuclear magnetization. Nevertheless, some decay mechanisms for nuclear singlet order persist. One such mechanism is called scalar relaxation of the second kind (SR2K) and involves the relaxation of additional nuclei ("third spins") which have scalar couplings to the spin-1/2 pair. This mechanism requires a difference between the couplings of at least one third spin with the two members of the spin-1/2 pair, and depends on the longitudinal relaxation time of the third spin. The SR2K mechanism of nuclear singlet relaxation has previously been examined in the case where the relaxation rate of the additional spins is on the time scale of the nuclear Larmor frequency. In this paper, we consider a different regime, in which the longitudinal relaxation of the third spins is on a similar time scale to the J-coupling between the members of the spin pair. This regime is often encountered when the spin-1/2 pair has scalar couplings to nearby deuterium nuclei. We show that the SR2K mechanism may be suppressed in this regime by applying a radiofrequency field which is resonant either with the members of the spin pair, or with the third spins. These phenomena are analyzed theoretically and by numerical simulations, and demonstrated experimentally on a diester of [13C2, 2H2]-labeled fumarate in solution.

Recent advances in ruthenium-based olefin metathesis.

Ruthenium-based olefin metathesis catalysts, known for their functional group tolerance and broad applicability in organic synthesis and polymer science, continue to evolve as an enabling technology in these areas. A discussion of recent mechanistic investigations is followed by an overview of selected applications.

Pol I transcription and pre-rRNA processing are coordinated in a transcription-dependent manner in mammalian cells.

Pre-rRNA synthesis and processing are key steps in ribosome biogenesis. Although recent evidence in yeast suggests that these two processes are coupled, the nature of their association is unclear. In this report, we analyze the coordination between rDNA transcription and pre-rRNA processing in mammalian cells. We found that pol I transcription factor UBF interacts with pre-rRNA processing factors as analyzed by immunoprecipitations, and the association depends on active rRNA synthesis. In addition, injections of plasmids containing the human rDNA promoter and varying lengths of 18S rDNA into HeLa nuclei show that pol I transcription machinery can be recruited to rDNA promoters regardless of the product that is transcribed, whereas subgroups of pre-rRNA processing factors are recruited to plasmids only when specific pre-rRNA fragments are produced. Our observations suggest a model for sequential recruitment of pol I transcription factors and pre-rRNA processing factors to elongating pre-rRNA on an as-needed basis rather than corecruitment to sites of active transcription.

Perinucleolar compartment prevalence has an independent prognostic value for breast cancer.

The perinucleolar compartment (PNC) is a multicomponent nuclear structure enriched with RNAs transcribed by RNA pol III and RNA binding proteins. Studies in cultured cells showed an association between PNC and transformed phenotype. To evaluate the relationship between structure and malignancy in vivo, we examined PNC prevalence (the percentage of cells containing at least one PNC) in normal and cancerous paraffin-embedded breast tissues using immunohistochemistry against a PNC-associated protein. Five hundred nuclei in the most active area of each sample were scored for PNC prevalence. The results show that PNC prevalence significantly correlates with the progression of breast cancer (by the criteria of staging). PNC prevalence in primary tumors, lymph nodes, and distant metastases shows a stepwise increase from a median of 23% in primary tumors to approximately 100% in distant metastases. In addition, univariate and multivariate (controlling for tumor size and grade) analyses show that early-stage patients with invasive ductal carcinomas containing a higher PNC prevalence have a significantly poorer prognosis. These findings link PNC prevalence with the progression of breast cancer in vivo and suggest that PNC-containing cells have metastatic advantages. These findings also show the potential of PNC prevalence as a prognostic marker for breast cancer.

Comparison of pulsatile ocular blood flow in Indians and Europeans.

PURPOSE: To compare pulsatile ocular blood flow (POBF) in Europeans and Indians and provide reference values for a group of healthy Indians. PATIENTS AND METHODS: Measurement with the POBF Tonograph was performed on healthy Indian subjects in India (n=252). A further 80 subjects (40 of Indian descent and 40 Europeans) underwent measurements in Cambridge, England. The instrument used for measurement was the same for both the studies. RESULTS: The mean POBF in the Indians in India was found to be 1176 microl/min. The mean POBF value in the Europeans was found to be 1033 microl/min and that for Indians in England was 1061 microl/min. The difference between the POBF within groups was significant (one-way ANOVA P<0.05) with the POBF of Indians in India being higher than Europeans and Indians in the UK. The difference between the Europeans and Indians in the UK did not reach statistical significance. CONCLUSIONS: POBF values in Indians living in India were found to be considerably higher than the previously published normal value of 650 microl/min in European studies and other studies for other racial groups. The reason for this apparent difference may be instrument-related rather than genetic because such a large difference was not observed when a comparison was performed in the UK. In addition, the results for both groups in our comparative study were still considerably higher than reported in previous studies. The POBF of Indians in India is slightly higher than the POBF of people of Indian ethnic origin in England.

Effect of corneal parameters on measurements using the pulsatile ocular blood flow tonograph and Goldmann applanation tonometer.

AIMS: To investigate the effect of central corneal thickness and corneal curvature on intraocular pressure measurements using the pulsatile ocular blood flow tonograph and the Goldmann applanation tonometer, and to assess the agreement between the pulsatile ocular blood flow tonograph and the Goldmann applanation tonometer in intraocular pressure measurement. METHODS: 479 subjects underwent intraocular pressure measurements with the Goldmann applanation tonometer and the pulsatile ocular blood flow tonograph. Of these, 334 patients underwent additional measurement of central corneal thickness with an ultrasonic pachymeter and corneal curvature measurement with a keratometer. RESULTS: The intraocular pressure measurements obtained with both the Goldmann applanation tonometer and the pulsatile ocular blood flow tonograph varied with central corneal thickness and mean keratometric reading. Intraocular pressure measured using the Goldmann applanation tonometer increased by 0.027 mm Hg per micro m increase in central corneal thickness. Intraocular pressure measured using the pulsatile ocular blood flow tonograph increased by 0.048 mm Hg per micro m increase in central corneal thickness. For an increase of 1 mm of mean corneal curvature there was rise in intraocular pressure of 1.14 mm Hg measured by the Goldmann applanation tonometer and of 2.6 mm Hg measured by the pulsatile ocular blood flow tonograph. When compared to the Goldmann applanation tonometer, the pulsatile ocular blood flow tonograph underestimated at low intraocular pressure and overestimated at higher intraocular pressure. CONCLUSION: Central corneal thickness and corneal curvature affected measurements obtained with the pulsatile ocular blood flow tonograph more than they affected measurements obtained with the Goldmann applanation tonometer.

Nucleocytoplasmic shuttling of polypyrimidine tract-binding protein is uncoupled from RNA export.

Polypyrimidine tract binding protein, PTB/hnRNP I, is involved in pre-mRNA processing in the nucleus and RNA localization and translation in the cytoplasm. In this report, we demonstrate that PTB shuttles between the nucleus and cytoplasm in an energy-dependent manner. Deletion mutagenesis demonstrated that a minimum of the N terminus and RNA recognition motifs (RRMs) 1 and 2 are necessary for nucleocytoplasmic shuttling. Deletion of RRM3 and 4, domains that are primarily responsible for RNA binding, accelerated the nucleocytoplasmic shuttling of PTB. Inhibition of transcription directed by either RNA polymerase II alone or all RNA polymerases yielded similar results. In contrast, selective inhibition of RNA polymerase I did not influence the shuttling kinetics of PTB. Furthermore, the intranuclear mobility of GFP-PTB, as measured by fluorescence recovery after photobleaching analyses, increased significantly in transcriptionally inactive cells compared with transcriptionally active cells. These observations demonstrate that nuclear RNA transcription and export are not necessary for the shuttling of PTB. In addition, binding to nascent RNAs transcribed by RNA polymerase II and/or III retards both the nuclear export and nucleoplasmic movement of PTB. The uncoupling of PTB shuttling and RNA export suggests that the nucleocytoplasmic shuttling of PTB may also play a regulatory role for its functions in the nucleus and cytoplasm.

Regulation of ribosome biogenesis within the nucleolus.

Ribosome biogenesis is both necessary for cellular adaptation, growth, and proliferation as well as a major energetic and biosynthetic demand upon cells. For these reasons, ribosome biogenesis requires precise regulation to balance supply and demand. The complexity of ribosome biogenesis gives rise to many steps and opportunities where regulation could take place. For trans-acting factors involved in ribosome biogenesis in the nucleolus, there may be a dynamic coordination, both spatially and temporally, that regulates their functions from the transcription of rDNA to the assembly and export of preribosomal particles. Here we summarize most of the described regulations on ribosome biogenesis in the nucleolus. However, these may represent only a small fraction of a larger picture. Further studies are required to determine the initial signals, signal transduction pathways utilized, and the specific targets of these regulatory modifications and how these are used to control ribosome biogenesis as a whole.

Facility of accommodation in myopia.

PURPOSE: Myopes have been shown to have abnormal accommodative characteristics. This study investigated the characteristics of accommodation facility in myopic and emmetropic students. METHODS: Distance and near positive and negative accommodation response time components of facility were measured over a 1 min period using a -2.00 D/zero lens pair for distance responses and a +/-2.00 D lens pair for near responses. 79 students (37 myopes and 42 emmetropes) aged 18-27 years acted as subjects. Subjects were masked, and the results were analysed in a masked fashion. RESULTS: Mean distance facility was significantly lower (9.7 cycles per minute (cpm)) in the myopic group compared with the mean distance facility in the emmetropic group (15.6 cpm; p < 0.005). There was no significant difference in the near facilities of the two groups (11.5 cpm in myopes vs 12.9 cpm in emmetropes). Positive accommodation response time for distance vision was greater than 4 s in 45% of myopes and in 9% of emmetropes. CONCLUSIONS: Our findings confirm that myopes tend to have abnormal accommodation responses to blur. Distance facility, but not near facility of accommodation is more frequently reduced in myopes than in emmetropes.

Ring-closing metathesis of olefinic peptides: design, synthesis, and structural characterization of macrocyclic helical peptides.

Heptapeptides containing residues with terminal olefin-derivatized side chains (3 and 4) have been treated with ruthenium alkylidene 1 and undergone facile ring-closing olefin metathesis (RCM) to give 21- and 23-membered macrocyclic peptides (5 and 6). The primary structures of peptides 3 and 4 were based upon a previously studied heptapeptide (2), which was shown to adopt a predominantly 3(10)-helical conformation in CDCl(3) solution and an alpha-helical conformation in the solid state. Circular dichroism, IR, and solution-phase (1)H NMR studies strongly suggested that acyclic precursors 3 and 4 and the fully saturated macrocyclic products 7 and 8 also adopted helical conformations in apolar organic solvents. Single-crystal X-ray diffraction of cyclic peptide 8 showed it to exist as a right-handed 3(10)-helix up to the fifth residue. Solution-phase NMR structures of both acyclic peptide 4 and cyclic peptide 8 in CD(2)Cl(2) indicated that the acyclic diene assumes a loosely 3(10)-helical conformation, which is considerably rigidified upon macrocyclization. The relative ease of introducing carbon-carbon bonds into peptide secondary structures by RCM and the predicted metabolic stability of these bonds renders olefin metathesis an exceptional methodology for the synthesis of rigidified peptide architectures.

Scalar coupling across the hydrogen bond in 1,3- and 1,4-diols.

[reaction: see text] Scalar coupling between hydroxyl groups sharing a 1,4- or 1, 3-intramolecular hydrogen bond can be readily detected using standard 2D NMR techniques. Analyses of isotopically perturbed 2D J-resolved spectra have revealed (h2)J to be on the order of 0.3 Hz in systems such as diols 2 and 3. A standard 2D COSY experiment, modified to detect small coupling constants, is shown to be a convenient method for detecting (h2)J in these systems. Couplings derived from density functional finite perturbation theory (DFT/FPT) are found to be in general agreement with experimental data.

Free-space accommodative response and minus lens-induced accommodative response in pre-school children.

PURPOSE: The purpose of this study was to investigate and compare the accommodative accuracy of pre-school children between free-space accommodative stimulus and minus lens-induced accommodative stimulus. METHODS: The accommodative responses of 59 pre-school children, ages 2 to 5 years, were measured objectively with a Canon Autoref R-1 under two different conditions, giving an optical accommodative stimulus of 1 D; first, viewing a target placed 100 cm away (free-space stimulus), and second, viewing a distant target through a -1.00 D lens. RESULTS: The mean lag of the accommodative response for the free-space accommodative stimulus is 0.24 D, while the mean lag of the accommodative response with the minus lens-induced accommodative stimulus is 0.69 D. The lag of accommodative response for free-space accommodative stimulus is significantly lower than the lag of accommodative response for minus lenses. CONCLUSIONS: Pre-school children can accommodate more accurately toward a free-space accommodative stimulus, as compared with minus lens-induced accommodative stimulus.

Corneal response to orthokeratology.

PURPOSE: The technique of orthokeratology produces a corneal response to the mechanical pressures exerted by rigid contact lenses. This paper reports a study which investigated the topographic and pachometric corneal changes induced by orthokeratology. METHODS: Six young myopic subjects (11 eyes) wore "accelerated orthokeratology" lenses (OK-74; Contex Inc., Sherman Oaks, CA) in a high Dk material (AirPerm; Dk = 88) for 28 days. Corneal and epithelial thickness were measured topographically using the Holden-Payor optical micropachometer, and corneal topography was monitored using the EyeSys system. RESULTS: Refractive error change reached 1.71 +/- 0.59 D reduction in myopia after 28 days. After 1 day of lens wear, statistically significant central corneal flattening was noted, which progressed to reach 0.22 +/- 0.07 mm (1.19 +/- 0.38 D) at 28 days. A trend toward central epithelial thinning was apparent, reaching statistical significance on day 28 (7.1 +/- 7.1 microm; 9.6%). Midperipheral corneal thickening was also found approximately 2.5 mm from the corneal center, which was statistically significant by day 14 (13.0 +/- 11.1 microm; 2.4%). Calculations using Munnerlyn's formula indicate that changes in corneal sagittal height based on topographical thickness changes across the flattened central 5.25-mm zone can account for the refractive changes observed. CONCLUSIONS: These findings suggest that the initial corneal response to orthokeratology may be explained by redistribution of corneal tissue, rather than by overall bending of the cornea.

Size and number of epithelial cells washed from the cornea after contact lens wear.

Using the technique of Fullard and Wilson, tear samples were collected atraumatically from the corneas of 14 rigid gas permeable (RGP) contact lens wearers, 14 soft contact lens (SCL) wearers, and 22 controls who did not wear contact lenses. Samples were analyzed for the number, size, and contiguity of the cellular contents. Both RGP and SCL groups had, on average, significantly fewer cells per sample than controls (median values were 11 and 12 cells vs. 44). The difference remained significant even after a correction of three cells was applied to account for cells adherent to the lenses. Approximately 80% of epithelial cells from both lens wear and control eyes were found in isolation, whereas 20% were contiguous with 1 or 2 other cells. This result is compatible with the majority of epithelial cells having exfoliated from the surface of the cornea after detaching from their neighbors, rather than detaching in sheets of cells. The cell length and cell area were analyzed. RGP wearers had significantly smaller cells than controls, whereas SCL wearers had slightly (but significantly) larger cells. These results may reflect changes in the exfoliation mechanism of the corneal epithelium in contact lens wear.

Nutritional requirements of the corneal epithelium and anterior stroma: clinical findings.

PURPOSE: To monitor the health of the epithelium and the anterior stroma when porous membranes are implanted into the feline cornea and to determine membrane diffusivity characteristics needed to maintain corneal integrity. METHODS: Filtration membranes in a range of effective pore sizes of less than 15 nm (groups 1 and 2, n = 11), 25 nm (group 3, n = 8), 50 nm (group 4, n = 16), and 100 nm (group 5, n = 15) were implanted into an interlamellar corneal pocket of the stroma. The implanted membranes ranged in thickness from 6 nm to 15 nm and were between 8 mm and 12 mm in diameter. Animals were monitored for clinical signs of intolerance to the implants. RESULTS: At 1 month, thinning and ulceration had occurred in the epithelium and the anterior stroma of all animals in groups 1 and 2; epithelial changes, anterior stromal thinning, and ulceration had developed in 75% of animals of group 3; 50% of animals showed vascularization and only 7% showed epithelial degeneration in group 4; and local anterior stromal thinning was observed in 7% of animals in group 5, indicating clinical acceptance of the implanted membrane. In the long term (greater than 50 days), 30% and 73% of the group 4 and 5 corneas, respectively, were clinically quiet. Analysis of the failure times indicated an inverse relation between failure rate and pore size: less than 15 nm > 25 nm > 50 nm > 100 nm. The difference between the 100-nm and 50-nm membranes was significant (P = 0.03). CONCLUSIONS: A corneal implant must have a porosity greater than that provided by 50-nm membranes. The 100-nm membranes used in this study establish the porosity needed to satisfy the nutritional requirements of the cornea.

Changes in the interpupillary distance with age and its effect on the near fusion free position.

PURPOSE: The effect of the rate of the interpupillary distance (IPD) change with age on the near fusion free position was investigated in the present study. METHODS: Three hundred and fifty-two children aged from 2 to 16 years of age were tested. Interpupillary distance was estimated with a modified Viktorin's method and the near fusion free position was measured with a modified Maddox Wing. RESULTS: The results reveal a significant difference in IPD growth rate between females and males and between children aged below and above 5 years. Most children below 5 years of age were orthophoric, but heterophoria became more common above 5 years of age. CONCLUSION: The present study suggests that the oculomotor control system for convergence can compensate for age changes in IPD.

Optometric findings in velocardiofacial syndrome.

Velocardiofacial syndrome (VCFS) is a congenital genetically determined condition with skeletal, heart, and facial anomalies. Almost all children have learning problems, despite an often normal or above average IQ. We have examined 10 children with a prior diagnosis of VCFS. All were mildly to moderately hyperopic. Most had poor accommodation, and convergence was clinically poor. About 30% had tortuous retinal vessels. VCFS may be detected in the optometry clinic, and optometric management may ameliorate the learning problems in some cases.

Incision depth affects the recovery of corneal sensitivity and neural regeneration in the cat.

To assess the effect of incision depth on the recovery of corneal sensitivity and neural regeneration, adult domestic cats underwent either 8-mm circular nonpenetrating keratotomies or penetrating autografts. The contralateral eye served as control. Corneal sensitivity was determined at various intervals after surgery. The depth of incision in the nonpenetrating keratotomies, assessed by optical pachometry, ranged between 49 and 91% of total corneal thickness. The animals were ranked based on the depth of incision and the average sensitivity within the keratotomy over the 1-yr recovery period. A significant negative correlation was found between incision depth and the recovery of sensitivity (Spearman rank-order correlation r = -0.84, P less than 0.05). Some recovery of sensitivity was found in the center and periphery of the incised zone when incision depth was less than 53% of total corneal thickness. With deeper incisions, the center of the incised zone remained insensitive throughout the measurement period, while the periphery of the incised zone showed a slight recovery of corneal sensitivity, proportional to incision depth. The recovery of corneal sensitivity was higher in a small annular region just distal to the incision site. When incision depth exceeded approximately 53%, gold chloride impregnation showed that the resultant reinnervation was confined to single intraepithelial axons or localized regions of irregular epithelial fibers. With shallower incisions, the deeper stromal trunks were spared, resulting in the persistence of a reduced subepithelial plexus and basal epithelial leashes. We have shown that when an incision severs all stromal trunks, the neural regeneration is insufficient for functional recovery of corneal sensitivity at the center of an 8-mm keratotomy.(ABSTRACT TRUNCATED AT 250 WORDS)