Development of an sRNA-mediated conditional knockdown system for .

We describe a new protein depletion method that uses an engineered small RNA (sRNA) to inhibit translation of a target gene. In proof-of-principle experiments, we induced functional knockdown of IncA, a fusion-mediating inclusion membrane protein, as shown with Western blots, loss of IncA staining at the inclusion membrane, and production of multiple chlamydial inclusions within an infected cell. These effects were titratable and reversible.

Chlamydia trachomatis induces disassembly of the primary cilium to promote the intracellular infection.

Chlamydia trachomatis is a clinically important bacterium that infects epithelial cells of the genitourinary and respiratory tracts and the eye. These differentiated cells are in a quiescent growth state and have a surface organelle called a primary cilium, but the standard Chlamydia cell culture infection model uses cycling cells that lack primary cilia. To investigate if these differences are relevant, we performed infections with host cells that have a primary cilium.

A Reverse Genetic Approach for Studying sRNAs in Chlamydia trachomatis.

sRNAs are noncoding transcripts that play critical roles in posttranscriptional regulation in prokaryotes. In the intracellular bacterium Chlamydia, sRNAs have been identified, but functional studies have been limited to an E. coli heterologous system.

Differential Effects of Small Molecule Inhibitors on the Intracellular Chlamydia Infection.

Chlamydia are obligate intracellular bacteria that reside within a membrane-bound compartment called the chlamydial inclusion inside a eukaryotic host cell. These pathogens have a complex biphasic developmental cycle, which involves conversion between a replicating, but noninfectious, reticulate body (RB) and an infectious elementary body (EB). Small molecule inhibitors have been reported to have deleterious effects on the intracellular Chlamydia infection, but these studies have typically been limited in terms of assays and time points of analysis.

Chlamydia and HPV induce centrosome amplification in the host cell through additive mechanisms.

Based on epidemiology studies, Chlamydia trachomatis has been proposed as a co-factor for human papillomavirus (HPV) in the development of cervical cancer. These two intracellular pathogens have been independently reported to induce the production of extra centrosomes, or centrosome amplification, which is a hallmark of cancer cells. We developed a cell culture model to systematically measure the individual and combined effects of Chlamydia and HPV on the centrosome in the same host cell.

The Small Molecule H89 Inhibits Inclusion Growth and Production of Infectious Progeny.

is an obligate intracellular bacterium and the most common reportable cause of human infection in the United States. This pathogen proliferates inside a eukaryotic host cell, where it resides within a membrane-bound compartment called the chlamydial inclusion. It has an unusual developmental cycle, marked by conversion between a replicating form, the reticulate body (RB), and an infectious form, the elementary body (EB).

Stochastic Chlamydia Dynamics and Optimal Spread.

Chlamydia trachomatis is an important bacterial pathogen that has an unusual developmental switch from a dividing form (reticulate body or RB) to an infectious form (elementary body or EB). RBs replicate by binary fission within an infected host cell, but there is a delay before RBs convert into EBs for spread to a new host cell. We developed stochastic optimal control models of the Chlamydia developmental cycle to examine factors that control the number of EBs produced.

Restoring spinopelvic harmony with lateral lumbar interbody fusion: is it a realistic goal?

Background: The importance of spinopelvic harmony [pelvic incidence (PI) = lumbar lordosis (LL) ±10 degrees] is well established in the literature. We aimed to determine whether lateral lumbar interbody fusion (LLIF) surgery in isolation is successful in restoring spinopelvic harmony, and whether the surgery maintained the relationship in those who present in a balanced state.

Methods: A retrospective radiographic analysis was performed on patients who underwent LLIF surgery, followed by posterior instrumented fusion, between January 2012 to August 2019 by a single surgeon (AD).

Aristotle's dream: Evolutionary and neural aspects of aesthetic communication in the arts.

Art in general perception is something that transcends our notion of reality. In view of the earliest findings in Paleolithic sites, their abstract appearance and sometimes ceremonial context increased their status of a secret language. Even the first figurative cave paintings remained in a context of an encoded semantic whole.

Function of Golgi-centrosome proximity in RPE-1 cells.

The close physical proximity between the Golgi and the centrosome is a unique feature of mammalian cells that has baffled scientists for years. Several knockdown and overexpression studies have linked the spatial relationship between these two organelles to the control of directional protein transport, directional migration, ciliogenesis and mitotic entry. However, most of these conditions have not only separated these two organelles, but also caused extensive fragmentation of the Golgi, making it difficult to dissect the specific contribution of Golgi-centrosome proximity.

Culture by Nature. Familial roots of ambivalent human social behavior and its cultural extensions in large-scale societies. A contribution of Human and Cultural Ethology.

Since Hobbes' dictum of "homo homini lupus" in 17 century the tenor in European philosophy was the one of human nature in need of strong external rules to restrain from mutual aggression, inter-personal and inter-group. Behavioral measures and rules limiting a presumably aggressive human nature were perceived as purely moral and cultural. Evolutionary theory in the 19 century seemed to confirm the notion of a self-interested individual, at least in parts.

Relationship between sagittal balance and adjacent segment disease in surgical treatment of degenerative lumbar spine disease: meta-analysis and implications for choice of fusion technique.

Study Design: Meta-analysis.

Objective: To conduct a meta-analysis investigating the relationship between spinopelvic alignment parameters and development of adjacent level disease (ALD) following lumbar fusion for degenerative disease. ALD is a degenerative pathology that develops at mobile segments above or below fused spinal segments.

Replication-dependent size reduction precedes differentiation in Chlamydia trachomatis.

Chlamydia trachomatis is the most common cause of bacterial sexually transmitted infection. It produces an unusual intracellular infection in which a vegetative form, called the reticulate body (RB), replicates and then converts into an elementary body (EB), which is the infectious form. Here we use quantitative three-dimensional electron microscopy (3D EM) to show that C.

Reconstruction of Thoracic Spine Using a Personalized 3D-Printed Vertebral Body in Adolescent with T9 Primary Bone Tumor.

Background: Neurosurgery and spine surgery have the potential to benefit from the use of 3-dimensional printing (3DP) technology due to complex anatomic considerations and the delicate nature of surrounding structures. We report a procedure that uses a 3D-printed titanium T9 vertebral body implant post T9 vertebrectomy for a primary bone tumor.

Case Description: A 14-year-old female presented with progressive kyphoscoliosis and a pathologic fracture of the T9 vertebra with sagittal and coronal deformity due to a destructive primary bone tumor.

Spatial analysis of Cdc42 activity reveals a role for plasma membrane-associated Cdc42 in centrosome regulation.

The ability of the small GTPase Cdc42 to regulate diverse cellular processes depends on tight spatial control of its activity. Cdc42 function is best understood at the plasma membrane (PM), where it regulates cytoskeletal organization and cell polarization. Active Cdc42 has also been detected at the Golgi, but its role and regulation at this organelle are only partially understood.

The daughter centriole controls ciliogenesis by regulating Neurl-4 localization at the centrosome.

The two centrioles of the centrosome differ in age and function. Although the mother centriole mediates most centrosome-dependent processes, the role of the daughter remains poorly understood. A recent study has implicated the daughter centriole in centriole amplification in multiciliated cells, but its contribution to primary ciliogenesis is unclear.

Range of motion, sacral screw and rod strain in long posterior spinal constructs: a biomechanical comparison between S2 alar iliac screws with traditional fixation strategies.

Background: S1 screw failure and L5/S1 non-union are issues with long fusions to S1. Improved construct stiffness and S1 screw offloading can help avoid this. S2AI screws have shown to provide similar stiffness to iliac screws when added to L3-S1 constructs.

The importance of loading the periphery of the vertebral endplate.

Background: Commercial fusion cages typically provide support in the central region of the endplate, failing to utilize the increased compressive strength around the periphery. This study demonstrates the increase in compressive strength that can be achieved if the bony periphery of the endplate is loaded.

Methods: Sixteen cadaveric lumbar vertebrae (L1-L5) were randomly divided into two even groups.

Spine surgery in Nepal: the 2015 earthquake.

At noon on Saturday, 25 April 2015, a 7.8 magnitude earthquake struck Nepal. It was centered in the Himalaya northwest of Kathmandu, the capital of over 1 million people.

Induction and inhibition of CPAF activity during analysis of Chlamydia-infected cells.

Studies of the chlamydial protease CPAF have been complicated by difficulties in distinguishing bona fide intracellular proteolysis from in vitro proteolysis. This confounding issue has been attributed to CPAF activity in lysates from Chlamydia-infected cells. We compared three methods that have been used to inhibit in vitro CPAF-mediated proteolysis: (1) pre-treatment of infected cells with the inhibitor clasto-lactacystin, (2) direct cell lysis in 8 M urea and (3) direct lysis in hot 1% SDS buffer.

Art as behaviour--an ethological approach to visual and verbal art, music and architecture.

In recent years, the fine arts, architecture, music and literature have increasingly been examined from the vantage point of human ethology and evolutionary psychology. In 2011 the authors formed the research group 'Ethology of the Arts' concentrating on the evolution and biology of perception and behaviour. These novel approaches aim at a better understanding of the various facets represented by the arts by taking into focus possible phylogenetic adaptations, which have shaped the artistic capacities of our ancestors.

Signaling at the Golgi during mitosis.

The Golgi complex of mammalian cells is composed of interconnected stacks of flattened cisternae that form a continuous membrane system in the pericentriolar region of the cell. At the onset of mitosis, this so-called Golgi ribbon is converted into small tubular-vesicular clusters in a tightly regulated fragmentation process, which leads to a temporary loss of the physical Golgi-centrosome proximity. Mitotic Golgi breakdown is required for Golgi partitioning into the two daughter cells, cell cycle progression and may contribute to the dispersal of Golgi-associated signaling molecules.

Par6γ is at the mother centriole and controls centrosomal protein composition through a Par6α-dependent pathway.

The centrosome contains two centrioles that differ in age, protein composition and function. This non-membrane bound organelle is known to regulate microtubule organization in dividing cells and ciliogenesis in quiescent cells. These specific roles depend on protein appendages at the older, or mother, centriole.