Chimeric CRISPR-CasX enzymes and guide RNAs for improved genome editing activity.

A compact protein with a size of <1,000 amino acids, the CRISPR-associated protein CasX is a fundamentally distinct RNA-guided nuclease when compared to Cas9 and Cas12a. Although it can induce RNA-guided genome editing in mammalian cells, the activity of CasX is less robust than that of the widely used S. pyogenes Cas9. Here, we show that structural features of two CasX homologs and their guide RNAs affect the R-loop complex assembly and DNA cleavage activity. Cryo-EM-based structural engineering of either the CasX protein or the guide RNA produced two new CasX genome editors (DpbCasX-R3-v2 and PlmCasX-R1-v2) with significantly improved DNA manipulation efficacy. These results advance both the mechanistic understanding of CasX and its application as a genome-editing tool.

Phosphine Catalysis of the Fluorination of Unactivated Tertiary Alkyl Chlorides under Mild and Convenient Conditions.

Due to the significance of organofluorine compounds in disciplines ranging from medicine to agriculture to materials science, the invention of new methods for the creation of carbon-fluorine bonds is an important objective. Among the underdeveloped dimensions in this area are the fluorination of hindered alkyl halides (particularly chlorides) and the discovery of catalysts for such fluorination processes. Herein, we report a mild method for the fluorination of unactivated tertiary alkyl chlorides (and bromides), catalyzed by inexpensive PPh3. This straightforward process is compatible with a range of hindered electrophiles and a variety of functional groups.

Diffusion prepared pseudo-continuous arterial spin labeling reveals blood-brain barrier dysfunction in patients with CADASIL.

OBJECTIVES: Diffusion prepared pseudo-continuous arterial spin labeling (DP-pCASL) is a newly proposed MRI method to noninvasively measure the function of the blood-brain barrier (BBB). We aim to investigate whether the water exchange rate across the BBB, estimated with DP-pCASL, is changed in patients with cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL), and to analyze the association between the BBB water exchange rate and MRI/clinical features of these patients. METHODS: Forty-one patients with CADASIL and thirty-six age- and sex-matched controls were scanned with DP-pCASL MRI to estimate the BBB water exchange rate (kw). The MRI lesion burden, the modified Rankin scale (mRS), and the neuropsychological scales were also examined. The association between kw and MRI/clinical features was analyzed. RESULTS: Compared with that in the controls, kw in patients with CADASIL was decreased at normal-appearing white matter (NAWM) (t = - 4.742, p < 0.001), cortical gray matter (t = - 5.137, p < 0.001), and deep gray matter (t = - 3.552, p = 0.001). After adjustment for age, gender, and arterial transit time, kw at NAWM was negatively associated with the volume of white matter hyperintensities (ß = - 0.754, p = 0.001), whereas decreased kw at NAWM was independently associated with an increased risk of abnormal mRS scale (OR = 1.058, 95% CI: 1.013-1.106, p = 0.011) in these patients. CONCLUSIONS: This study found that the BBB water exchange rate was decreased in patients with CADASIL. The decreased BBB water exchange rate was associated with an increased MRI lesion burden and functional dependence of the patients, suggesting the involvement of BBB dysfunction in the pathogenesis of CADASIL. CLINICAL RELEVANCE STATEMENT: DP-pCASL reveals BBB dysfunction in patients with CADASIL. The decreased BBB water exchange rate is associated with MRI lesion burden and functional dependence, indicating the potential of DP-pCASL as an evaluation method for disease severity. KEY POINTS: • DP-pCASL reveals blood-brain barrier dysfunction in patients with CADASIL. • Decreased BBB water exchange rate, an indicator of BBB dysfunction detected by DP-pCASL, was associated with MRI/clinical features of patients with CADASIL. • DP-pCASL can be used as an evaluation method to assess the severity of disease in patients with CADASIL.

AllenDigger, a Tool for Spatial Expression Data Visualization, Spatial Heterogeneity Delineation, and Single-Cell Registration Based on the Allen Brain Atlas.

Spatial transcriptomics can be used to capture cellular spatial organization and has facilitated new insights into different biological contexts, including developmental biology, cancer, and neuroscience. However, its wide application is still hindered by its technical challenges and immature data analysis methods. Allen Brain Atlas (ABA) provides a great source for spatial gene expression throughout the mouse brain at various developmental stages with in situ hybridization image data. To the best of our knowledge, the portal developed to access spatial expression data is not very useful to biologists. Here, we developed a toolkit to collect and preprocess expression data from the ABA and allow a friendlier query to visualize the spatial distribution of genes of interest, characterize the spatial heterogeneity of the brain, and register cells from single-cell transcriptomics data to fine anatomical brain regions via machine learning methods with high accuracy. AllenDigger will be very helpful to the community in precise spatial gene expression queries and add extra spatial information to further interpret the scRNA-seq data in a cost-effective manner.

The transition from invariant to action-dependent visual object representation in human dorsal pathway.

The human brain can efficiently process action-related visual information, which supports our ability to quickly understand and learn others' actions. The visual information of goal-directed action is extensively represented in the parietal and frontal cortex, but how actions and goal-objects are represented within this neural network is not fully understood. Specifically, which part of this dorsal network represents the identity of goal-objects? Is such goal-object information encoded at an abstract level or highly interactive with action representations? Here, we used functional magnetic resonance imaging with a large number of participants (n = 94) to investigate the neural representation of goal-objects and actions when participants viewed goal-directed action videos. Our results showed that the goal-directed action information could be decoded across much of the dorsal pathway, but in contrast, the invariant goal-object information independent of action was mainly localized in the early stage of dorsal pathway in parietal cortex rather than the down-stream areas of the parieto-frontal cortex. These results help us to understand the relationship between action and goal-object representations in the dorsal pathway, and the evolution of interactive representation of goal-objects and actions along the dorsal pathway during goal-directed action observation.

Enantioselective Synthesis of cis-Decalins by Merging the Birch Reduction and Inverse-Electron-Demand Diels-Alder Reaction.

Herein, we show that the combination of the Birch reduction of readily available anisole derivatives and the catalytic asymmetric inverse-electron-demand Diels-Alder reaction of 2-pyrones can serve as a powerful platform for the diverse synthesis of synthetically important cis-decalin scaffolds. Enabled by a well-modified chiral bis(oxazoline) ligand/CuII complex, a wide range of polysubstituted cis-decalin scaffolds with up to six contiguous stereocenters were generated efficiently. The synthetic potential of this method is demonstrated by the concise synthesis of the sesquiterpene (+)-occidentalol and a key intermediate for seven triterpenes. Mechanistic studies suggest the 1,3-cyclohexadienes formed in situ are the key intermediates, and efficient kinetic resolution occurs when C2- and/or C3-substituted 1,4-cyclohexadienes are utilized as substrates. DFT calculations elucidated that the Diels-Alder reaction proceeds in a stepwise fashion and revealed the origins of the stereoselectivities.

Multi-echo balanced SSFP with a sequential phase-encoding order for functional MR imaging at 7T.

PURPOSE: To develop a 2D multi-echo passband balanced SSFP (bSSFP) sequence using an echo-train readout with a sequential phase-encoding order (sequential multi-echo bSSFP), and evaluate its performance in fast functional brain imaging at 7 T. METHODS: As images of sequential multi-echo bSSFP exhibit multiple ghosts due to periodic k-space modulations, a GRAPPA-based reconstruction method was proposed to eliminate ghosting artifacts. MRI experiments were performed to compare the image quality of multi-echo bSSFP and conventional single-echo bSSFP. Submillimeter-resolution fMRI using a checkerboard visual stimulus was conducted to compare the activation characteristics of multi-echo bSSFP, conventional single-echo bSSFP and standard gradient-echo EPI (GE-EPI). RESULTS: A higher mean structural similarity index was found between images of single-echo bSSFP and multi-echo bSSFP with a shorter echo train length (ETL). Multi-echo bSSFP (ETL = 3) showed higher temporal SNR (tSNR) values than GRAPPA-accelerated single-echo bSSFP (R = 2). In submillimeter-resolution fMRI experiments, multi-echo bSSFP (ETL = 3) approached the imaging speed of GRAPPA-accelerated single-echo bSSFP (R = 2), but without tSNR penalty and reduced activation due to acceleration. The median t-value and the number of significantly activated voxels were comparable between GE-EPI and multi-echo bSSFP (ETL = 3) that provides virtually distortion-free functional images and inherits the activation patterns of conventional bSSFP. CONCLUSION: Sequential multi-echo bSSFP (ETL = 3) is suitable for fast fMRI with submillimeter in-plane resolution, and offers an option to accelerate bSSFP imaging without tSNR penalty like parallel imaging.

New imaging features of tuberous sclerosis complex: A 7 T MRI study.

Few in vivo studies have focused on the perivenous association of tubers and iron deposition in the deep gray nuclei in patients with tuberous sclerosis complex (TSC). We investigated this possible relationship in TSC patients using susceptibility weighted imaging (SWI) at 7 T. SWI with high spatial resolution and enhanced sensitivity was performed on 11 TSC patients in comparison with 15 age- and sex-matched healthy controls. The relationship between tubers and veins was evaluated. In addition, the phase images of SWI were processed to produce local field shift (LFS) maps to quantify iron deposition. The mean LFS in the deep gray nuclei was compared between the TSC patients and healthy controls using a covariance analysis. Venous involvement was observed in 211 of the 231 (91.3%) cortical tubers on SWI. The slender tubers often oriented around the long axis of penetrating veins, possibly because cortical tubers typically developed and/or migrated along venous vasculatures. A significant difference in LFS of the thalamus was detected between the TSC patients and healthy controls (3.36 ± 0.50 versus 3.01 ± 0.39, p < 0.01). The new in vivo imaging features observed at 7 T provide valuable insights into the possible venous association of TSC lesions and iron accumulation in the deep gray nuclei. Our results may lead to a better understanding of the pathological changes involved in TSC under in vivo conditions.

Topological dominance in peripheral vision.

The question of what peripheral vision is good for, especially in pattern recognition, is one of the most important and controversial issues in cognitive science. In a series of experiments, we provide substantial evidence that observers' behavioral performance in the periphery is consistently superior to central vision for topological change detection, while nontopological change detection deteriorates with increasing eccentricity. These experiments generalize the topological account of object perception in the periphery to different kinds of topological changes (i.e., including introduction, disappearance, and change in number of holes) in comparison with a broad spectrum of geometric properties (e.g., luminance, similarity, spatial frequency, perimeter, and shape of the contour). Moreover, when the stimuli were scaled according to cortical magnification factor and the task difficulty was well controlled by adjusting luminance of the background, the advantage of topological change detection in the periphery remained. The observed advantage of topological change detection in the periphery supports the view that the topological definition of objects provides a coherent account for object perception in peripheral vision, allowing pattern recognition with limited acuity.

Topological change induces an interference effect in visual working memory.

The "irrelevant-change distracting effect" refers to the effect of changes in irrelevant features on the performance of the target feature, which has frequently been used to study information processing in visual working memory (VWM). In the current study, we reported a novel interference effect in VWM: the topological-change interference effect (TCIE). In a series of six experiments, we examined the influence of topological and nontopological changes as irrelevant features on VWM using a color change detection paradigm. The results revealed that only topological changes, although task irrelevant, could produce a significant interference effect. In contrast, nontopological changes did not produce any evident interference effect. Moreover, the TCIE was a stable and lasting effect, regardless of changes in locations, reporting methods, particular stimulus figures, the other salient feature dimensions and delay interval times. Therefore, our results support the notion that topological invariance that defines perceptual objects plays an essential role in maintaining representations in VWM.

Robust functional mapping of layer-selective responses in human lateral geniculate nucleus with high-resolution 7T fMRI.

The lateral geniculate nucleus (LGN) of the thalamus is the major subcortical relay of retinal input to the visual cortex. It plays important roles in visual perception and cognition and is closely related with several eye diseases and brain disorders. Primate LGNs mainly consist of six layers of monocular neurons with distinct cell types and functions. The non-invasive measure of layer-selective activities of the human LGN would have broad scientific and clinical implications. Using high-resolution functional magnetic resonance imaging (fMRI) at 7 Tesla (T) and carefully designed visual stimuli, we achieved robust functional mapping of eye-specific and also magnocellular/parvocellular-specific laminar patterns of the human LGN. These laminar patterns were highly reproducible with different pulse sequences scanned on separate days, between different subjects, and were in remarkable consistency with the simulation from high-resolution histology of the human LGNs. These findings clearly demonstrate that 7T fMRI can robustly resolve layer-specific responses of the human LGN. This paves the way for future investigation of the critical roles of the LGN in human visual perception and cognition, as well as the neural mechanisms of many developmental and neurodegenerative diseases.

Improved depiction of subthalamic nucleus and globus pallidus internus with optimized high-resolution quantitative susceptibility mapping at 7 T.

The subthalamic nucleus (STN) and globus pallidus internus (GPi) are commonly used targets in deep-brain stimulation (DBS) surgery for the treatment of movement disorders. The success of DBS critically depends on the spatial precision of stimulation. By taking advantage of good contrast between iron-rich deep-brain nuclei and surrounding tissues, quantitative susceptibility mapping (QSM) has shown promise in differentiating the STN and GPi from the adjacent substantia nigra and globus pallidus externus, respectively. Nonlinear morphology-enabled dipole inversion (NMEDI) is a widely used QSM algorithm, but the image quality of reconstructed susceptibility maps relies on the regularization parameter selection. To date, few studies have systematically optimized the regularization parameter at the ultra-high field of 7 T. In this study, we optimized the regularization parameter in NMEDI to improve the depiction of STN and GPi at different spatial resolutions at both 3 T and 7 T. The optimized QSM images were further compared with other susceptibility-based images, including T2*-weighted (T2*w), R2*, susceptibility-weighted, and phase images. QSM showed better depiction of deep-brain nuclei with clearer boundaries compared with the other methods, and 7 T QSM at 0.35 × 0.35 × 1.0 mm3 demonstrated superior performance to the others. Our findings suggest that optimized high-resolution QSM at 7 T allows for improved delineation of deep-brain nuclei with clear and sharp borders between nuclei, which may become a promising tool for DBS nucleus preoperative localization.

Stereoselective On-Surface Cyclodehydrofluorization of a Tetraphenylporphyrin and Homochiral Self-Assembly.

The thermally induced cyclodehydrofluorization of iron tetrakis(pentafluorophenyl)porphyrin proceeded highly stereoselectively to give a prochiral product on a gold surface in an ultrahigh vacuum, whereas dehydrocyclization of the respective iron tetrakisphenylporphyrin did not show such selectivity. Stereoselectivity was predominantly observed for closely packed layers, which is an indication of intermolecular cooperativity and steric constraints induced by adjacent species. Density functional theory identified intermolecular packing constraints as the origin of such selectivity during the reaction. Scanning tunneling microscopy revealed the formation of an enantiomerically pure two-dimensional self-assembly as a conglomerate of mirror domains. On-surface two-dimensional topochemistry, as reported herein, may open new routes for stereoselective synthesis.

Visualization of the lenticulostriate arteries at 3T using black-blood T1-weighted intracranial vessel wall imaging: comparison with 7T TOF-MRA.

OBJECTIVES: The objective of this study was to explore the feasibility of using intracranial T1-weighted vessel wall imaging (VWI) to visualize the lenticulostriate arteries (LSAs) at 3T. MATERIAL AND METHODS: Thirteen healthy volunteers were examined with VWI at 3T and TOF-MRA at 7T during the same day. On the vascular skeletons obtained by manual tracing, the number of stems and branches of LSAs were counted. On the most prominent branch in every hemisphere, the contrast-to-noise ratio (CNR), the full length and the local length (5-15 mm above MCAs) were measured and compared between the two methods. Nine stroke patients with intracranial artery stenosis were also recruited into the study. The branches of LSAs were compared between the symptomatic and asymptomatic side. RESULTS: The extracted vascular trees were in good agreement between 7T TOF-MRA and 3T VWI. The two acquisitions showed similar numbers of the LSA stems. The number of branches revealed by 3T VWI was slightly lower than 7T TOF. The full lengths were slightly lower by VWI at 3T (p = 0.011, ICC = 0.917). The measured local lengths (5-15 mm from MCAs) showed high coherence between VWI and TOF-MRA (p = 0.098, ICC = 0.970). In stroke patients, 12 plaques were identified on MCA segments, and nine plaques were located on the symptomatic side. The average numbers of LSA visualized by 3T VWI were 4.3±1.3 on the symptomatic side and 5.0±1.1 on the asymptomatic side. CONCLUSION: 3T VWI is capable of depicting LSAs, particularly the stems and the proximal segments, with comparable image quality to that of 7T TOF-MRA. KEY POINTS: • T1-weighted intracranial VWI at 3T allows for black-blood MR angiography of lenticulostriate artery. • 3T intracranial VWI depicts the stems and proximal segments of the lenticulostriate arteries comparable to 7T TOF-MRA. • It is feasible to assess both large vessel wall lesions and lenticulostriate vasculopathy in one scan.

Visual working memory representation as a topological defined perceptual object.

The question of what the basic unit is of visual working memory remains one of the most fundamental and controversial issues. In the current study, we proposed a unique perspective based on early topological perception to describe the nature of representation in visual working memory. In a series of updating change-detection tasks, the repetition-benefit effect on color memory was not affected when items in the second memory array underwent massive changes of nontopological features from the first memory array. However, when the topological properties of an item changed, the repetition-benefit effect was destroyed, suggesting that the item was perceived as a new object impairing the original memory. Hence, our results suggest that a perceptual object defined by its topological invariance might be a unique perspective from which to describe representations of visual working memory.

Noncontrast-enhanced time-resolved 4D dynamic intracranial MR angiography at 7T: A feasibility study.

BACKGROUND: Arterial spin labeling (ASL) based-noncontrast-enhanced 4D MR angiography (NCE 4D MRA) shows potential in characterizing cerebrovascular hemodynamics in cerebrovascular disorders. Ultrahigh-field theoretically benefits ASL signal with increased inherent signal-to-noise ratio (SNR) and prolonged blood T1 , which may provide improved delineation of vasculature in 4D MRA. PURPOSE: To investigate the feasibility of NCE 4D MRA using 3D Cartesian trajectory and stack-of-stars (SOS) golden angle radial trajectory at 7T. STUDY TYPE: A prospective study. SUBJECTS: Six normal volunteers and eight patients with arteriovenous malformation (AVM). FIELD STRENGTH/SEQUENCE: NCE 4D MRA with Cartesian and radial trajectories were performed at 3T and 7T. ASSESSMENT: Subjective image quality of 4D MRA was evaluated using a 4-point scale by two experienced neuroradiologists. The characterization of AVM components with 4D MRA and DSA was also graded using the Spetzler-Martin grading scale. STATISTICAL TESTS: Cohen's kappa coefficient was calculated to evaluate the agreement between two readers within each 4D MRA technique (Cartesian and Radial). A Wilcoxon signed-rank test was performed to compare the subjective image quality scores of 4D MRA between Cartesian and radial trajectories, and between 7T and 3T, respectively. RESULTS: Good-to-excellent image quality was achieved in 4D MRA with both Cartesian (3.83 ± 0.41) and radial (3.42 ± 0.49) acquisitions in healthy volunteers at 7T. However, markedly reduced scan time was needed with radial acquisition. 4D MRA at 7T (3.31 ± 0.59) shows better delineation of AVM lesion features, especially the vein drainage, compared with that of 3T (2.83 ± 0.75), although no statistical significance was achieved (P = 0.180). DATA CONCLUSION: The feasibility of ASL based 4D MRA at 7T with Cartesian and SOS golden angle radial acquisition was demonstrated. The clinical evaluation of 4D MRA in AVMs between 3T and 7T suggested 7T 4D MRA images acquired with radial acquisition demonstrate excellent delineation of AVM features, especially the draining veins. LEVEL OF EVIDENCE: 2 Technical Efficacy Stage 2 J. Magn. Reson. Imaging 2017.

Direct localisation of the human pedunculopontine nucleus using MRI: a coordinate and fibre-tracking study.

OBJECTIVES: To image the pedunculopontine tegmental nucleus (PPN), a deep brain stimulation (DBS) target for Parkinson disease, using MRI with validated results. METHODS: This study used the MP2RAGE sequence with high resolution and enhanced grey-white matter contrast on a 7-T ultra-high-field MRI system to image the PPN as well as a diffusion spectrum imaging method on a 3-T MRI system to reconstruct the main fibre systems surrounding the PPN. The coordinates of the rostral and caudal PPN poles of both sides were measured in relation to the third and fourth ventricular landmarks on the 7-T image. RESULTS: The boundary of the PPN was delineated, and showed morphology consistent with previous histological works. The main fibres around the PPN were reconstructed. The pole coordinate results combined with the fibre spatial relationships validate the imaging results. CONCLUSIONS: A practical protocol is provided to directly localise the PPN using MRI; the position and morphology of the PPN can be obtained and validated by locating its poles relative to two ventricular landmarks and by inspecting its spatial relationship with the surrounding fibre systems. This technique can be potentially used in clinics to define the boundary of the PPN before DBS surgery for treatment of Parkinson disease in a more precise and reliable manner. KEY POINTS: • Combined information helps localise the PPN as a DBS target for PD patients • Scan the PPN at 7 T and measure its coordinates against different ventricular landmarks • Reconstruct the main fibres around the PPN using diffusion spectrum imaging.

Improved visualization of superficial temporal artery using segmented time-of-flight MR angiography with venous suppression at 7T.

PURPOSE: To evaluate the quality of time-of-flight MR angiography (TOF-MRA) with venous suppression at 7T on imaging superficial temporal artery (STA). METHODS: A recently developed segmented TOF technique with reduced specific absorption rate (SAR) of venous suppression (VS) module was employed to achieve high-resolution arterial angiography without the contamination of venous signal. Images of segmented TOF with VS at 7T, TOF without VS at 7T, and TOF with VS at 3T were collected on 17 healthy volunteers. The number of STA branches and their local contrast achieved by the three methods were quantified and compared using paired t test. RESULTS: Segmented TOF with VS at 7T successfully suppressed venous signal without reducing the contrast of arterial angiography. The numbers of STA branches in 7T images were significantly higher than that in 3T images (5.79 vs. 4.50, p < 0.001). The contrast of 7T segmented TOF was significantly higher than 3T TOF (7.21 vs. 5.56, p = 0.006). CONCLUSION: Segmented TOF with VS at 7T displayed more branches of STA, while eliminating the signal of superficial temporal vein (STV). The improved visualization of STA will potentially facilitate the pre-operative assessment of STA in STA-MCA bypass surgery.

Magnetic resonance imaging of tuberous sclerosis complex with or without epilepsy at 7 T.

PURPOSE: This study was conducted to determine the benefit of magnetic resonance imaging (MRI) at 7 T in detecting structural lesions and previously unidentified abnormalities in patients with tuberous sclerosis complex (TSC). METHODS: Thirteen patients with TSC (8-36 years, seven males) previously diagnosed by 3 T MRI underwent additional imaging at 7 T, which included T1-weighted magnetization-prepared rapid gradient-echo (MPRAGE), T2-weighted turbo spin echo (TSE), SPACE fluid attenuated inversion recovery (FLAIR), susceptibility weighted imaging (SWI), white matter suppressed (WM-suppressed), and gray-white matter tissue border enhancement (GW-TBE) MPRAGE sequences. Subtle lesions, tuberal morphology, and perituberal cortex abnormalities were examined and compared to those observed at 3 T MRI using standard sequences. RESULTS: Improved visualization of TSC lesions was achieved in all subjects at 7 T compared to 3 T imaging, and three subjects received resective surgery. The 7 T T1- and T2-weighted images had high spatial resolution and provided a clear delineation of the perituberal cortex. SWI revealed abnormal blood vessel morphology. WM-suppressed and GW-TBE protocols, adjusted specifically for TSC imaging, aided in visualizing lesions and uncovered more extensive subtle lesions and abnormalities beyond the conventionally detected tubers. CONCLUSIONS: Due to its high spatial resolution and powerful new imaging protocols, 7 T MRI provided a better evaluation of subtle lesions and perituberal details compared with conventional MRI at 3 T, with potential implications for diagnosis and operative treatment of TSC and its related epilepsy.

Association of Dose and Frequency on the Survival of Patients on Maintenance of Hemodialysis in China: A Kaplan-Meier and Cox-Proportional Hazard Model Analysis.

BACKGROUND Dialysis frequency and dose are controversial prognostic factors of hemodialysis morbidity and mortality. The aim of this study was to find out the effect of frequency and dosage of dialysis on mortality and survival in a group of Chinese hemodialysis patients. MATERIAL AND METHODS In total, 183 patients seen from February 2008 to January 2018, who were on maintenance hemodialysis for at least 3 months, were included in the study cohort. An anonymized database of age, gender, diabetic status, comorbidities, date of initiation of dialysis, hematological characters, biochemical variables, and status of survived or died was established from DICOM (Digital Imaging and Communications in Medicine) files of patients. Kaplan-Meier and Cox-proportional hazard model was used for calculation of survival over time at 95% confidence level. RESULTS Overall, the 10-year survival rate was 27%. Kaplan-Meier analysis showed patient survival as 94% at one-year, 59% at 5-years, and 27% at 10-years. Hemoglobin, serum albumin, calcium, potassium, phosphorous, calcium-phosphorous-products, and hemodialysis frequency and the dose had a significant effect on survival. Cox regression proportional hazard model showed that patients with serum albumin level of >4 g/dL were better associated with survival. Patients who underwent twice-weekly hemodialysis had 4.26 times less chance of survival as compared to patients with thrice-weekly hemodialysis. A higher dialysis dose of >1.2 spKt/V offered better survival as compared to a lower dose of <1.2 spKt/V. CONCLUSIONS Hypoalbuminemia, hemodialysis time, and hemodialysis frequency were significantly associated with mortality.