Nanoscale resolution of microbial fiber degradation in action

The lives of microbes unfold at the micron scale, and their molecular machineries operate at the nanoscale. Their study at these resolutions is key toward achieving a better understanding of their ecology. We focus on cellulose degradation of the canonical Clostridium thermocellum system to comprehend how microbes build and use their cellulosomal machinery at these nanometer scales. Degradation of cellulose, the most abundant organic polymer on Earth, is instrumental to the global carbon cycle. We reveal that bacterial cells form 'cellulosome capsules' driven by catalytic product-dependent dynamics, which can increase the rate of hydrolysis. Biosynthesis of this energetically costly machinery and cell growth are decoupled at the single-cell level, hinting at a division-of-labor strategy through phenotypic heterogeneity. This novel observation highlights intrapopulation interactions as key to understanding rates of fiber degradation

Insight into the functional organization of nuclear lamins in health and disease

Lamins are the main component of the nuclear lamina, a protein meshwork at the inner nuclear membrane which primarily provide mechanical stability to the nucleus. Lamins, type V intermediate filament proteins, are also involved in many nuclear activities. Structural analysis of nuclei revealed that lamins form 3.5nm thick filaments often interact with nuclear pore complexes. Mutations in the LMNA gene, encoding A-type lamins, have been associated with at least 15 distinct diseases collectively termed laminopathies, including muscle, metabolic and neurological disorders, and premature aging syndrome. It is unclear how laminopathic mutations lead to such a wide array of diseases, essentially affecting almost all tissues

Unveiling the polarity of actin filaments by cryo-electron tomography

The actin cytoskeleton plays a fundamental role in numerous cellular processes, such as cell motility, cytokinesis, and adhesion to the extracellular matrix. Revealing the polarity of individual actin filaments in intact cells would foster an unprecedented understanding of cytoskeletal processes and their associated mechanical forces. Cryo-electron tomography provides the means for high-resolution structural imaging of cells. However, the low signal-to-noise ratio of cryo-tomograms obscures the high frequencies, and therefore the polarity of actin filaments cannot be directly measured. Here, we developed a method that enables us to determine the polarity of actin filaments in cellular cryo-tomograms. We applied it to reveal the actin polarity distribution in focal adhesions, and show a linear relation between actin polarity and distance from the apical boundary of the adhesion site

A lamin A/C variant causing striated muscle disease provides insights into filament organization

The LMNA gene encodes the A-type lamins, which polymerize into ∼3.5-nm-thick filaments and, together with B-type lamins and associated proteins, form the nuclear lamina. Mutations in LMNA cause a wide variety of pathologies. In this study, we analyzed the nuclear lamina of embryonic fibroblasts from LmnaH222P/H222P mice, which develop cardiomyopathy and muscular dystrophy. Although the organization of the lamina appeared unaltered, there were changes in chromatin and B-type lamin expression. An increase in nuclear size and consequently a relative reduction in heterochromatin near the lamina allowed for a higher resolution structural analysis of lamin filaments using cryo-electron tomography. This was most apparent when visualizing lamin filaments in situ and using a nuclear extraction protocol. Averaging of individual segments of filaments in LmnaH222P/H222P mouse fibroblasts resolved two polymers that constitute the mature filaments. Our findings provide better views of the organization of lamin filaments and the effect of a striated muscle disease-causing mutation on nuclear structure

Computational analyses reveal spatial relationships between nuclear pore complexes and specific lamins

Nuclear lamin isoforms form fibrous meshworks associated with nuclear pore complexes (NPCs). Using datasets prepared from subpixel and segmentation analyses of 3D-structured illumination microscopy images of WT and lamin isoform knockout mouse embryo fibroblasts, we determined with high precision the spatial association of NPCs with specific lamin isoform fibers. These relationships are retained in the enlarged lamin meshworks of Lmna-/- and Lmnb1-/- fibroblast nuclei. Cryo-ET observations reveal that the lamin filaments composing the fibers contact the nucleoplasmic ring of NPCs. Knockdown of the ring-associated nucleoporin ELYS induces NPC clusters that exclude lamin A/C fibers but include LB1 and LB2 fibers. Knockdown of the nucleoporin TPR or NUP153 alters the arrangement of lamin fibers and NPCs. Evidence that the number of NPCs is regulated by specific lamin isoforms is presented. Overall the results demonstrate that lamin isoforms and nucleoporins act together to maintain the normal organization of lamin meshworks and NPCs within the nuclear envelope

CITROBACTER PERITONITIS: TWO CASES AND REVIEW OF THE LITERATURE

One of the uncommon gram negative organisms causing peritonitis in peritoneal dialysis patients is Citrobacter. Because of this organism's resistant nature, treatment for Citrobacter peritonitis may be difficult, and removal of the catheter may be necessary in refractory cases. Here we present 2 cases of peritonitis caused by this organism and fully treated with antibiotics. The literature contains only a limited number of reports on Citrobacter peritonitis, mostly case reports or portions of general papers about the microbiological spectrum of peritonitis in the relevant units. Until enough data about this micro-organism have been accumulated to map out an approach, it is wise to individualize treatment by watching the response of the patient during the wait for the antibiogram result and not to hesitate to remove the catheter if the clinical situation deteriorates

Placement of Hemodialysis Catheters with a Technical, Functional, and Anatomical Viewpoint

Aims. Vascular access is of prime importance for hemodialysis patients. We aimed to study early complications of hemodialysis catheters placed in different central veins in patients with acute or chronic renal failure with or without ultrasound (US ) guidance. Material and Methods. Patients who were admitted to our unit between March 2008 and December 2010 with need for vascular access have been included. 908 patients were examined for their demographic parameters, primary renal disease, and indication for catheterization, type and location of the catheter, implantation technique, and acute complications. Results. The mean age of the patients was 60.6 ± 16.0 years. 643 (70.8 %) of the catheters were temporary while 265 (29.2%) were permanent. 684 catheters were inserted to internal jugular veins, 213 to femoral, and 11 to subclavian veins. Arterial puncture occurred in 88 (9.7%) among which 13 had resultant subcutaneous hematoma. No patient had lung trauma and there had been no need for removal of the catheter or a surgical intervention for complications. US guidance in jugular vein and experience of operator decreased arterial puncture rate. Conclusion. US-guided replacement of catheter to internal jugular vein would decrease complication rate. Referral to invasive nephrologists may decrease use of subclavian vein. Experience improves complication rates even under US guidance

Placement of Hemodialysis Catheters with a Technical, Functional, and Anatomical Viewpoint

Aims. Vascular access is of prime importance for hemodialysis patients. We aimed to study early complications of hemodialysis catheters placed in different central veins in patients with acute or chronic renal failure with or without ultrasound (US) guidance. Material and Methods. Patients who were admitted to our unit between March 2008 and December 2010 with need for vascular access have been included. 908 patients were examined for their demographic parameters, primary renal disease, and indication for catheterization, type and location of the catheter, implantation technique, and acute complications. Results. The mean age of the patients was 60.6 +/- 16.0 years. 643 (70.8 %) of the catheters were temporary while 265 (29.2%) were permanent. 684 catheters were inserted to internal jugular veins, 213 to femoral, and 11 to subclavian veins. Arterial puncture occurred in 88 (9.7%) among which 13 had resultant subcutaneous hematoma. No patient had lung trauma and there had been no need for removal of the catheter or a surgical intervention for complications. US guidance in jugular vein and experience of operator decreased arterial puncture rate. Conclusion. US-guided replacement of catheter to internal jugular vein would decrease complication rate. Referral to invasive nephrologists may decrease use of subclavian vein. Experience improves complication rates even under US guidance

A Case of Wegener's Granulomatosis Simulating Lung Cancer and Treatment with Plasmapheresis

Wegener's Granulomatosis (WG) is a rare disease characterized by granulomatous vasculitis of the upper and lower respiratory tract, kidneys and other organs. Herein, we present a case analyzed with an initial diagnosis of metastatic lung cancer, but later, as the patient was diagnosed with WG, she was treated with plasmapheresis. A 62-year-old female was admitted with fatigue, cough, dyspnea and hemoptysis. She was examined in a hospital specialized in chest diseases with initial diagnosis of metastatic lung cancer due to her symptoms for the last six months; she was referred to our clinic due to uremia. On physical examination, the patient had tachypnea, orthopnea, pretibial edema and diffuse rales to the upper lung zones. Laboratory tests revealed urea of 211 mg/dl and creatinine of 6.3 mg/dl, besides leukocytosis and anemia. Based on the chest X-ray, which demonstrated nodular cavitating lesions, and on positive c-ANCA, the patient was diagnosed with WG and immunosuppressive treatment together with plasmapheresis was applied. The clinical and radiological findings resolved completely. Pulmonary involvement of WG is usually in the form of bilateral, cavitating, solitary nodule or mass with smooth borders, and may be confused with lung cancer, as in our case. The presence of alveolar hemorrhage increases mortality up to 50%. High-dose immunosuppressive therapy together with plasmapheresis may be lifesaving in these patients