Can occipital lobe bending, Gibraltar sign of superior sagittal sinus groove and jugular foramen dimensions predict transverse sinus dominance?

BACKGROUND: Asymmetry between the transverse sinuses (TS) is quite common. We sought to test the possible hypothesis that certain anatomical features - namely, occipital lobe bending, Gibraltar sign of superior sagittal sinus groove (SSS) and jugular foramen (JF) dimensions - can predict dominance of the transverse sinuses on routine axial T1- and T2-weighted images. MATERIALS AND METHODS: One hundred consecutively acquired combined MRI-MRV studies of brain were reviewed. On non-contrast axial T1WI, each reviewer assessed the occipital lobe bending, and Gibraltar sign of SSS groove; on axial T2-weighted images, JF dimensions were measured. TS cross-sectional area was measured on non-contrast sagittal 2-dimensional phase contrast MRV images and served as the reference standard. RESULTS: Of the 51 subjects with right-dominant TS, 37 had occipital bending to the right side and 35 showed sloping of the Gibraltar sign to right side. Of the 18 subjects with left dominant TS, 10 had occipital bending to left side and 13 showed left-sided sloping of the Gibraltar sign. Of the 31 subjects with co-dominant TS, 15 had no occipital bending and 20 showed no sloping of the Gibraltar sign. Mean right and left JF dimensions were higher in the right and left dominant TS respectively with no significant differences in patients with co-dominant sinus (p < 0.02). CONCLUSION: Right occipital bending had a good association with right TS dominance. The other two parameters- Gibraltar sign of superior sagittal sinus groove and jugular foramen dimensions - did not have a very good association with respect to TS dominance.

Altered cropping pattern and cultural continuation with declined prosperity following abrupt and extreme arid event at ~4,200 yrs BP: Evidence from an Indus archaeological site Khirsara, Gujarat, western India.

Archaeological sites hold important clues to complex climate-human relationships of the past. Human settlements in the peripheral zone of Indus culture (Gujarat, western India) are of considerable importance in the assessment of past monsoon-human-subsistence-culture relationships and their survival thresholds against climatic stress exerted by abrupt changes. During the mature phase of Harappan culture between ~4,600-3,900yrsBP, the ~4,100±100yrsBP time slice is widely recognized as one of the major, abrupt arid-events imprinted innumerous well-dated palaeo records. However, the veracity of this dry event has not been established from any archaeological site representing the Indus (Harappan) culture, and issues concerning timing, changes in subsistence pattern, and the likely causes of eventual abandonment (collapse) continue to be debated. Here we show a significant change in crop-pattern (from barley-wheat based agriculture to 'drought-resistant' millet-based crops) at ~4,200 yrs BP, based on abundant macrobotanical remains and C isotopes of soil organic matter (δ13CSOM) in an archaeological site at Khirsara, in the Gujarat state of western India. The crop-change appears to be intentional and was likely used as an adaptation measure in response to deteriorated monsoonal conditions. The ceramic and architectural remains of the site indicate that habitation survived and continued after the ~4,200yrsBP dry climatic phase, but with declined economic prosperity. Switching to millet-based crops initially helped inhabitants to avoid immediate collapse due to climatic stresses, but continued aridity and altered cropping pattern led to a decline in prosperity levels of inhabitants and eventual abandonment of the site at the end of the mature Harappan phase.

Combustion Synthesized Europium Doped LaAI11O18 Phosphors--An Electron Paramagnetic Resonance and Optical Study.

By adopting a facile solution combustion synthesis, crystallized europium doped blue phosphors were successfully prepared. These phosphors were characterized by X-ray diffraction (XRD), Fourier transform infrared absorption (FT-IR), energy dispersive analysis of X-ray, Electron Paramagnetic Resonance (EPR) and Photoluminescence (PL) experimental methods. The photoluminescence spectrum indicates Eu²âº and Eu³âº ions in these phosphors. The band at around 442 nm is attributed to the spin-allowed 4f65d¹ (2D) --> 4f7(8S7/2) transition of Eu²âº ions. The sharp bands at 591 (5D0 --> 7F1) and 616 (5D0 --> 7F2) nm are attributed to the spin-forbidden transitions of Eu³âº ions. The EPR spectra of as-prepared and post-treated LaAl11O18:Eu phosphors exhibit signals characteristic of La²âº and Eu²âº ions. The number of spins, Gibbs energy, magnetic susceptibility, Curie constant and effective magnetic moment values were calculated and compared at 296 and 110 K.

Receptor-mediated delivery of engineered nucleases for genome modification.

Engineered nucleases, which incise the genome at predetermined sites, have a number of laboratory and clinical applications. There is, however, a need for better methods for controlled intracellular delivery of nucleases. Here, we demonstrate a method for ligand-mediated delivery of zinc finger nucleases (ZFN) proteins using transferrin receptor-mediated endocytosis. Uptake is rapid and efficient in established mammalian cell lines and in primary cells, including mouse and human hematopoietic stem-progenitor cell populations. In contrast to cDNA expression, ZFN protein levels decline rapidly following internalization, affording better temporal control of nuclease activity. We show that transferrin-mediated ZFN uptake leads to site-specific in situ cleavage of the target locus. Additionally, despite the much shorter duration of ZFN activity, the efficiency of gene correction approaches that seen with cDNA-mediated expression. The approach is flexible and general, with the potential for extension to other targeting ligands and nuclease architectures.

Lamin A/C haploinsufficiency modulates the differentiation potential of mouse embryonic stem cells.

BACKGROUND: Lamins are structural proteins that are the major determinants of nuclear architecture and play important roles in various nuclear functions including gene regulation and cell differentiation. Mutations in the human lamin A gene cause a spectrum of genetic diseases that affect specific tissues. Most available mouse models for laminopathies recapitulate disease symptoms for muscle diseases and progerias. However, loss of human lamin A/C also has highly deleterious effects on fetal development. Hence it is important to understand the impact of lamin A/C expression levels on embryonic differentiation pathways. METHODOLOGY AND PRINCIPAL FINDINGS: We have investigated the differentiation potential of mouse embryonic stem cells containing reduced levels of lamin A/C by detailed lineage analysis of embryoid bodies derived from these cells by in vitro culture. We initially carried out a targeted disruption of one allele of the mouse lamin A/C gene (Lmna). Undifferentiated wild-type and Lmna(+/-) embryonic stem cells showed similar expression of pluripotency markers and cell cycle profiles. Upon spontaneous differentiation into embryoid bodies, markers for visceral endoderm such as α-fetoprotein were highly upregulated in haploinsufficient cells. However, neuronal markers such as ß-III tubulin and nestin were downregulated. Furthermore, we observed a reduction in the commitment of Lmna(+/-) cells into the myogenic lineage, but no discernible effects on cardiac, adipocyte or osteocyte lineages. In the next series of experiments, we derived embryonic stem cell clones expressing lamin A/C short hairpin RNA and examined their differentiation potential. These cells expressed pluripotency markers and, upon differentiation, the expression of lineage-specific markers was altered as observed with Lmna(+/-) embryonic stem cells. CONCLUSIONS: We have observed significant effects on embryonic stem cell differentiation to visceral endoderm, neuronal and myogenic lineages upon depletion of lamin A/C. Hence our results implicate lamin A/C level as an important determinant of lineage-specific differentiation during embryonic development.

Photophysical studies on the interaction of formamide and alkyl substituted amides with photoinduced electron transfer (PET) based acridinedione dyes in water.

Interaction of photoinduced electron transfer (PET) based acridinedione dye (ADR 1) with amides like formamide, acetamide and dimethylformamide (DMF) were investigated by fluorescence spectral techniques. A fluorescence enhancement accompanied with a blue shift in the emission maximum was observed on the addition of amides to ADR 1 dye, which possess C(6)H(4)(p-OCH(3)) in the 9th position of the basic acridinedione ring. The extent of fluorescence enhancement and the blue shift in the emission maximum of ADR 1 dye is of the order of DMF > acetamide > formamide. DMF, which is more hydrophobic and less polar, results in a higher extent of fluorescence enhancement and a larger shift in the emission maximum towards the blue region. On the addition of amides, the ADR 1 dye prefers to orient towards a more hydrophobic phase surrounded by more number of amide molecules. The fluorescence enhancement of ADR 1 dye is attributed to the suppression of PET process occurring through space. The influence of the hydrophobic nature and the polarity of the amides on the excited state properties of acridinedione dyes are elucidated by steady-state and time resolved fluorescence measurements.

LRRK2 expression in idiopathic and G2019S positive Parkinson's disease subjects: a morphological and quantitative study.

AIMS: Mutations in the gene encoding leucine-rich repeat kinase-2 (LRRK2) have been established as a common genetic cause of Parkinson's disease (PD). The distribution of LRRK2 mRNA and protein in the human brain has previously been described, although it has not been reported in PD cases with the common LRRK2 G2019S mutation. METHODS: To further elucidate the role of LRRK2 in PD, we determined the localization of LRRK2 mRNA and protein in post-mortem brain tissue from control, idiopathic PD (IPD) and G2019S positive PD cases. RESULTS: Widespread neuronal expression of LRRK2 mRNA and protein was recorded and no difference was observed in the morphological localization of LRRK2 mRNA or protein between control, IPD and G2019S positive PD cases. Using quantitative real-time polymerase chain reaction, we demonstrated that there is no regional variation in LRRK2 mRNA in normal human brain, but we have identified differential expression of LRRK2 mRNA with significant reductions recorded in limbic and neocortical regions of IPD cases compared with controls. Semi-quantitative analysis of LRRK2 immunohistochemical staining demonstrated regional variation in staining intensity, with weak LRRK2 immunoreactivity consistently recorded in the striatum and substantia nigra. No clear differences were identified in LRRK2 immunoreactivity between control, IPD and G2019S positive PD cases. LRRK2 protein was identified in a small proportion of Lewy bodies. CONCLUSIONS: Our data suggest that widespread dysregulation of LRRK2 mRNA expression may contribute to the pathogenesis of IPD.

Denaturation mechanism of BSA by urea derivatives: evidence for hydrogen-bonding mode from fluorescence tools.

Urea and alkyl urea derivatives, which posses a free N-H moiety in the urea molecular framework is responsible for the fluorescence quenching of BSA. Fluorescence quenching accompanied with a blue initially and subsequently a red shift in the emission maximum of BSA is observed on the addition of urea derivatives containing N-H moieties. On the contrary, a fluorescence enhancement accompanied with a shift in the emission maximum towards the blue region is observed on the addition of tetramethylurea (TMU). Urea derivatives, which posses a free N-H moiety acts as a perfect denaturant by direct hydrogen-bonding interaction with BSA resulting in the unfolding process. The unfolding of the buried tryptophan moieties to the aqueous phase does not occur, when all the N-H moieties in the urea are methyl substituted (TMU). Fluorescence spectral techniques reveal that the direct hydrogen-bonding interaction of the N-H moiety of urea molecular framework with the carbonyl oxygen moieties of BSA results in the unfolding of the tryptophan moieties to the aqueous phase, while that of the carbonyl oxygen of urea with the N-H moieties of BSA is definitely not involved in the denaturation process. Steady state and time-resolved fluorescence studies illustrate that the extent of protein folding occurs at a relatively lower concentration of unsymmetrical alkyl urea derivatives (butyl urea (BU) and ethyl urea (EU)), compared to that of urea.

Photophysical studies on the interaction of acridinedione dyes with universal protein denaturant: guanidine hydrochloride.

Photophysical studies of photoinduced electron transfer (PET) and non-PET based acridinedione dyes with guanidine hydrochloride (GuHCl) were carried out in water and methanol. Addition of GuHCl to photoinduced electron transfer (PET) based acridinedione dye (ADR 1) results in a fluorescence enhancement, whereas a non-PET based dye (ADR 2) shows no significant change in the fluorescence intensity and lifetime. Addition of GuHCl to ADR 1 dye in methanol results in single exponential decay behaviour, on the contrary a biexponential decay pattern was observed on the addition of GuHCl in water. Absorption and emission spectral studies of ADR 1 dye interaction with GuHCl reveals that the dye molecule is not in the protonated form in aqueous GuHCl solution, and the dye is confined to two distinguishable microenvironment in the aqueous phase. A large variation in the microenvironment around the dye molecule is created on the addition of GuHCl and this was ascertained by time-resolved area normalized emission spectroscopy (TRANES) and time-resolved emission spectroscopy (TRES). The dye molecule prefers to reside in the hydrophobic microenvironment, rather in the hydrophilic aqueous phase is well emphasized by time-resolved fluorescence lifetime studies. The mechanism of fluorescence enhancement of ADR 1 dye by GuHCl is attributed to the suppression of the PET process occurring through space.

Chromatin dynamics and gene positioning.

The mammalian cell nucleus provides a landscape where genes are regulated through their organization and association with freely diffusing proteins and nuclear domains. In many cases, specific genes are highly dynamic, and the principles governing their movements and interchromosomal interactions are currently under intensive study. Recent investigations have implicated actin and myosin in chromatin dynamics and gene expression. Here, we discuss our current understanding of the dynamics of the interphase genome and how it impacts nuclear organization and gene activity.

A genetic locus targeted to the nuclear periphery in living cells maintains its transcriptional competence.

The peripheral nuclear lamina, which is largely but not entirely associated with inactive chromatin, is considered to be an important determinant of nuclear structure and gene expression. We present here an inducible system to target a genetic locus to the nuclear lamina in living mammalian cells. Using three-dimensional time-lapse microscopy, we determined that targeting of the locus requires passage through mitosis. Once targeted, the locus remains anchored to the nuclear periphery in interphase as well as in daughter cells after passage through a subsequent mitosis. Upon transcriptional induction, components of the gene expression machinery are recruited to the targeted locus, and we visualized nascent transcripts at the nuclear periphery. The kinetics of transcriptional induction at the nuclear lamina is similar to that observed at an internal nuclear region. This new cell system provides a powerful approach to study the dynamics of gene function at the nuclear periphery in living cells.

PET suppression of acridinedione dyes by urea derivatives in water and methanol.

Spectroscopic investigations involving the interaction of acridinedione dyes with urea and its derivatives in water and methanol were carried out by absorption, steady-state fluorescence, and time-resolved fluorescence measurements. The hydrogen-bonding properties of urea and derivatives in aqueous solutions are found to be distinctly different from those observed in methanol. Urea, which can serve both as a hydrogen bond donor as well as an acceptor and has a unique hydrogen-bonding feature, helps in studying urea interaction with fluorophores in aqueous solutions, micelles, and alcohol. In our studies, we have used acridinedione dyes as the probe. We report that the hydrophobic interaction of urea with dye predominates by weakening of the hydrogen-bonding interaction of the solvent and urea derivatives with increase in the hydrophobicity of urea derivatives. In methanol, the hydrogen bonding between solvent and urea derivatives predominating over the hydrophobicity of the urea derivatives is observed. The presence of alkyl group substitution in the N-H moiety with a function of increasing concentration resulting in the creation of a more favorable hydrophobic environment to the dye molecule to reside in the hydrophobic shell phase rather than in the bulk aqueous phase is illustrated. The hydrophobic interaction of dye with urea in aqueous solution predominates because of the weakening of the hydrogen bonding of the solvent and urea derivatives, and the photoinduced electron transfer (PET) process is used as a marker to identify the hydrophobic interaction illustrated in our studies.

Lamin A/C speckles mediate spatial organization of splicing factor compartments and RNA polymerase II transcription.

The A-type lamins have been observed to colocalize with RNA splicing factors in speckles within the nucleus, in addition to their typical distribution at the nuclear periphery. To understand the functions of lamin speckles, the effects of transcriptional inhibitors known to modify RNA splicing factor compartments (SFCs) were examined. Treatment of HeLa cells with alpha-amanitin or 5,6-dichlorobenzimidazole riboside (DRB) inhibited RNA polymerase II (pol II) transcription and led to the enlargement of lamin speckles as well as SFCs. Removal of the reversible inhibitor DRB resulted in the reactivation of transcription and a rapid, synchronous redistribution of lamins and splicing factors to normal-sized speckles, indicating a close association between lamin speckles and SFCs. Conversely, the expression of NH2-terminally modified lamin A or C in HeLa cells brought about a loss of lamin speckles, depletion of SFCs, and down-regulation of pol II transcription without affecting the peripheral lamina. Our results suggest a unique role for lamin speckles in the spatial organization of RNA splicing factors and pol II transcription in the nucleus.

A general, norbornyl based approach to anti-Bredt alkenes via sequential RCM-fragmentation strategy.

A general protocol for the synthesis of bicyclo[n.3.1]frameworks with bridgehead double bond (anti-Bredt alkenes), from a common, readily available norbornyl precursor, involving sequential ring closure metathesis (RCM) and Wharton fragmentation is outlined.