Using a simplified two-dimensional divider-channel setup, we simulate the development process of red blood cell (RBC) flows in the entrance region of microvessels to study the wall shear stress (WSS) behaviors. Significant temporal and spatial variation in WSS is noticed. The maximum WSS magnitude and the strongest variation are observed at the channel inlet due to the close cell-wall contact. From the channel inlet, both the mean WSS and variation magnitude decrease, with a abrupt drop in the close vicinity near the inlet and then a slow relaxation over a relatively long distance; and a relative stable state with approximately constant mean and variation is established when the flow is well developed. The correlations between the WSS variation features and the cell free layer (CFL) structure are explored, and the effects of several hemodynamic parameters on the WSS variation are examined. In spite of the model limitations, the qualitative information revealed in this study could be useful for better understanding relevant processes and phenomena in the microcirculation.

Blood is a vital soft matter, and its normal circulation in the human body relies on the distribution of red blood cells (RBCs) at successive bifurcations. Understanding how RBCs are partitioned at bifurcations is key for the optimisation of microfluidic devices as well as for devising novel strategies for diagnosis and treatment of blood-related diseases. We report the dynamics of RBC suspensions flowing through a biomimetic vascular network incorporating three generations of microchannels and two classical types of bifurcations at the arteriole level. Our microfluidic experiments with dilute and semidilute RBC suspensions demonstrate the emergence of excessive heterogeneity of RBC concentration in downstream generations upon altering the network's outflow rates. Through parallel simulations using the immersed-boundary-lattice-Boltzmann method, we reveal that the heterogeneity is attributed to upstream perturbations in the cell-free layer (CFL) and lack of its recovery between consecutive bifurcations owing to suppressed hydrodynamic lift under reduced flow conditions. In the dilute/semidilute regime, this perturbation dominates over the effect of local fractional flow at the bifurcation and can lead to inherently unfavourable child branches that are deprived of RBCs even for equal flow split. Our work highlights the importance of CFL asymmetry cascading down a vascular network, which leads to biased phase separation that deviates from established empirical predictions.

OBJECTIVE

This study evaluates baseline patient characteristics and surgical parameters for risk factors of medical complications in ASD patients received posterior long level internal fixation.

METHODS

Analysis of consecutive patients who underwent posterior long-level instruction fixation for adult degenerative scoliosis (ADS) with a minimum of two year follow-up was performed. Pre-operative risk factors, intraoperative variables, peri-operative radiographic parameters, and surgical-related risk factors were collected to analyze the effect of risk factors on medical complications. Patients were separated into groups with and without medical complication. Then, complication group was further classified as major or minor medical complications. Potential risk factors were identified by univariate testing. Multivariate logistic regression was used to evaluate independent predictors of medical complications.

RESULTS

One hundred and thirty-one ADS patients who underwent posterior long segment pedicle screws fixation were included. Total medical complication incidence was 25.2%, which included infection (12.2%), neurological (11.5%), cardiopulmonary (7.6%), gastrointestinal (6.1%), and renal (1.5%) complications. Overall, 7.6% of patients developed major medical complications, and 17.6% of patients developed minor medical complications. The radiographic parameters of pre-operative and last follow-up had no significant difference between the groups of medical complications and the major or minor medical complications subgroups. However, the incidence of cerebrospinal fluid leak (CFL) in patients who without medical complications was much lower than that with medical complications (18.4 vs. 42.4%, P = 0.005). Independent risk factors for development of medical complications included smoking (OR = 6.45, P = 0.012), heart disease (OR = 10.07, P = 0.012), fusion level (OR = 2.12, P = 0.001), and length of hospital stay (LOS) (OR = 2.11, P = 0.000). Independent risk factors for development of major medical complications were diabetes (OR 6.81, P = 0.047) and heart disease (OR = 5.99, P = 0.049). Except for the last follow-up, Oswestry Disability Index and visual analog scale of the patient experienced medical complications trend higher score; the clinical outcomes have no significant difference between the medical and major complications groups.

CONCLUSIONS

Heart disease comorbidity is an independent risk factor for both medical and major medical complications. Smoking, fusion level, and LOS are independent risk factors for medical complication. Diabetes is the independent risk factors for major medical complications.

Sarcomeres, the fundamental contractile units of muscles, are conserved structures composed of actin thin filaments and myosin thick filaments. How sarcomeres are formed and maintained is not well understood. Here, we show that knockdown of Drosophila cofilin (DmCFL), an actin depolymerizing factor, disrupts both sarcomere structure and muscle function. The loss of DmCFL also results in the formation of sarcomeric protein aggregates and impairs sarcomere addition during growth. The activation of the proteasome delays muscle deterioration in our model. Furthermore, we investigate how a point mutation in CFLCFL function and leads to the muscle phenotypes observed in vivo. Our data provide significant insights to the role of CFLs during sarcomere formation, as well as mechanistic implications for disease progression in NM patients.

Keywords: Drosophila; actin depolymerizing factor; actin thin filaments; cofilin; muscle growth; nemaline myopathy; sarcomere addition; sarcomere homeostasis; sarcomeres; skeletal muscle.

Background: In the era of Option B+ and 'treat all' policies for HIV, challenges to retention in care are well documented. In Malawi, several large community-facility linkage (CFL) models have emerged to address these challenges, training lay health workers (LHW) to support the national prevention of mother-to-child transmission (PMTCT) programme. This qualitative study sought to examine how PMTCT LHW deployed by Malawi's three most prevalent CFL models respond to known barriers to access and retention to antiretroviral therapy (ART) and PMTCT.

Methods: We conducted a qualitative study, including 43 semi-structured interviews with PMTCT clients; 30 focus group discussions with Ministry of Health (MOH)-employed lay and professional providers and PMTCT LHWs; a facility CFL survey and 2-4 hours of onsite observation at each of 8 sites and in-depth interviews with 13 programme coordinators and MOH officials. Thematic analysis was used, combining inductive and deductive approaches.

Results: Across all three models, PMTCT LHWs carried out a number of 'targeted' activities that respond directly to a range of known barriers to ART uptake and retention. These include: (i) fulfilling counselling and educational functions that responded to women's fears and uncertainties; (ii) enhancing women's social connectedness and participation in their own care and (iii) strengthening service function by helping clinic-based providers carry out duties more efficiently and effectively. Beyond absorbing workload or improving efficiency, however, PMTCT LHWs supported uptake and retention through foundational but often intangible work to strengthen CFL, including via efforts to strengthen facility-side responsiveness, and build community members' recognition of and trust in services.

Conclusion: PMTCT LHWs in each of the CFL models examined, addressed social, cultural and health system factors influencing client access to, and engagement with, HIV care and treatment. Findings underscore the importance of person-centred design in the 'treat-all' era and the contribution LHWs can make to this, but foreground the challenges of achieving person-centredness in the context of an under-resourced health system. Further work to understand the governance and sustainability of these project-funded CFL models and LHW cadres is now urgently required.

Keywords: HIV; health services research; health systems; maternal health; public health.

Pretarsal, tarsal and tibial structures of the forelegs, midlegs and hindlegs of Pediculus humanus of humans and of Haematopinus apri Goureau, 1866 (Phthiraptera), a parasite of feral hogs, were studied using light microscopy and scanning electron microscopy. Details of the tibial thumb-like process (tl) with the spine of the thumb (spn), tarsal apophysis (ta) and the coupled finger-like process (cfl) can be observed in the leg photomicrograph of both species. A frontal view of the leg in open position shows the articulation of the claw: the structures of an open-closed system, a tooth row (te), rack-system (rs) and two telescopic columns (tc) which are present near the base of the claw in both species. In H. apri, we observed a pad-like structure, the flap-like tibial lobe (fl) on the ventral surface on the tarsus, the euplantulae, with several sensilla basiconica, which is present in each leg.

OBJECTIVE

To quantitatively evaluate the cartilage alteration of talus for chronic lateral ankle instability (LAI) with isolated anterior talofibular ligament (ATFL) tear and combined ATFL and calcaneofibular ligament (CFL) tear using T2 -mapping at 3.0T.

METHODS

In all, 27 patients including 17 with isolated ATFL tear and 10 with ATFL+CFL tear, and 21 healthy subjects were recruited. All participants underwent T2 -mapping scan at 3T and patients completed American Orthopaedic Foot and Ankle Society (AOFAS) scoring. The total talar cartilage (TTC) was segmented into six compartments: medial anterior (MA), medial center (MC), medial posterior (MP), lateral anterior (LA), lateral center (LC), and lateral posterior (LP). The T2 value of each compartment was measured from T2 -mapping images. Data were analyzed with one-way analysis of variance (ANOVA), Student's t-test, and Pearson's correlation coefficient.

RESULTS

The T2 values of MA, MC, MP, TTC in the ATFL group and MA, MC, MP, LC, LP, TTC in the ATFL+CFL group were higher than those in the control group (P < 0.05). Moreover, the T2 values of MC, MP, LC, and TTC in the ATFL+CFL group were higher than those in the ATFL group (P < 0.05). The T2 values of MA in both patient groups were negatively correlated with AOFAS scores (r = -0.596, r = -0.690, P < 0.05).

CONCLUSIONS

Chronic LAI with ATFL tear had a trend of increasing cartilage T2 values in talar trochlea, mainly involving medial cartilage compartments. Chronic LAI with ATFL+CFL tear might result in higher T2 values in a much larger cartilage region than with ATFL tear. MA could be the main cartilage compartment that may affect the patient's clinical symptoms.

METHODS

1 Technical Efficacy: Stage 3 J. Magn. Reson. Imaging 2018;47:69-77.

Accumulation of toxic protein assemblies and damaged mitochondria are key features of neurodegenerative diseases, which arise in large part from clearance defects in the Macroautophagy/autophagy-lysosome system. The autophagy cargo receptor SQSTM1/p62 plays a major role in the clearance of ubiquitinated cargo through Ser403 phosphorylation by multiple kinases. However, no phosphatase is known to physiologically dephosphorylate SQSTM1 on this activating residue. RNAi-mediated knockdown and overexpression experiments using genetically encoded fluorescent reporters and defined mutant constructs in cell lines, primary neurons, and brains show that SSH1, the canonical CFL (cofilin) phosphatase, mediates the dephosphorylation of phospho-Ser403-SQSTM1, thereby impairing SQSTM1 flux and phospho-MAPT/tau clearance. The inhibitory action of SSH1 on SQSTM1 is fully dependent on SQSTM1 Ser403 phosphorylation status and is separable from SSH1-mediated CFL activation. These findings reveal a unique action of SSH1 on SQSTM1 independent of CFL and implicate an inhibitory role of SSH1 in SQSTM1-mediated clearance of autophagic cargo, including phospho-MAPT/tau. Abbreviations: AAV: adeno-associated virus; Aβ42_O: amyloid β1-42 oligomers; AD: Alzheimer disease; CA3: cornu Ammonis 3; CSNK2/CK2: casein kinase 2; FCCP: 2-[2-[4-(trifluoromethoxy)phenyl]hydrazinylidene]-propanedinitrile; FTLD: frontotemporal lobar degeneration; GFP: green fluorescent protein; MAP1LC3/LC3: microtubule associated protein 1 light chain 3; SQSTM1/p62: sequestosome-1; PLA: proximity ligation assay; RFP: red fluorescent protein; RIPA: radioimmunoprecipitation assay; shRNA: short hairpin RNA; siRNA: small interfering RNA; Ser403: Serine403; SSH1: slingshot protein phosphatase 1; TBK1: TANK-binding kinase 1; ULK: unc-51 like kinase 1.

Keywords: Autophagy; CFL (cofilin); MAP1LC3/LC3; MAPT/tau; SQSTM1/p62; SSH1; mitochondria; tauopathy; ubiquitin.

We present a novel approach for analyzing biological time-series data using a context-free language (CFL) representation that allows the extraction and quantification of important features from the time-series. This representation results in Hierarchically AdaPtive (HAP) analysis, a suite of multiple complementary techniques that enable rapid analysis of data and does not require the user to set parameters. HAP analysis generates hierarchically organized parameter distributions that allow multi-scale components of the time-series to be quantified and includes a data analysis pipeline that applies recursive analyses to generate hierarchically organized results that extend traditional outcome measures such as pharmacokinetics and inter-pulse interval. Pulsicons, a novel text-based time-series representation also derived from the CFL approach, are introduced as an objective qualitative comparison nomenclature. We apply HAP to the analysis of 24 hours of frequently sampled pulsatile cortisol hormone data, which has known analysis challenges, from 14 healthy women. HAP analysis generated results in seconds and produced dozens of figures for each participant. The results quantify the observed qualitative features of cortisol data as a series of pulse clusters, each consisting of one or more embedded pulses, and identify two ultradian phenotypes in this dataset. HAP analysis is designed to be robust to individual differences and to missing data and may be applied to other pulsatile hormones. Future work can extend HAP analysis to other time-series data types, including oscillatory and other periodic physiological signals.

OBJECTIVE

To incorporate a diagnostic technique for measuring subtalar motion, namely "talar rotation", into the manual supination-anterior drawer stress radiographs for evaluation of the severity of rotational instability, and to determine its clinical relevance.

METHODS

Sixty-six patients with combined injuries of the anterior talofibular (ATFL) and calcaneofibular ligament (CFL) underwent three bilateral manual stress radiographs, and mean increments of anterior talar translation (mm), talar tilt (°), and talar rotation (%) in the injured ankle compared to the normal opposite side were measured with the technique. Intraobserver and interobserver reliability of each measure was assessed, and the difference in the degree of increments was compared according to the presence of additional cervical ligament insufficiency.

RESULTS

Ankle stress radiographic intraobserver and interobserver agreement was ICC = 0.91 and 0.82 for talar rotation (%), ICC = 0.64 and 0.51 for anterior talar translation, and ICC = 0.78 and 0.71 for talar tilt angle, respectively. In group 2 including patients with combined injuries of the ATFL and CFL along with additional cervical ligament insufficiency, a significantly higher increment of talar rotation, mean 6.4% (SD 3.4%), was observed compared to that of talar rotation, mean 4.1% (SD 2.7 ), in the other group (group 1) with an intact cervical ligament (p < 0.001).

CONCLUSIONS

A new comprehensive stress radiographic technique for diagnosis of chronic lateral ankle instability presented in this study might be a reliable and representable measurement tool to assess additional injury or instability of the subtalar joint.

METHODS

Prospective cohort study, Level II.

People with central field loss (CFL) lose information in the scotomatous region. Remapping is a method to modify images to present the missing information outside the scotoma. This study tested the hypothesis that remapping improves reading performance for subjects with simulated CFL.

Circular central scotomas, with diameters ranging from 4° to 16°, were simulated in normally sighted subjects using an eye tracker on either a head-mounted display (HMD) (experiments 1, 2) or a traditional monitor (experiment 3). In the three experiments, reading speed was measured for groups of 7, 11, and 13 subjects with and without remapping of text.

Remapping increased reading speed in all three experiments. On the traditional monitor, it increased reading speed by 34% (8°), 38% (12°), and 35% (16°). In the two HMD experiments, remapping increased reading speed only for the largest scotoma size, possibly due to latency of updating of the simulated scotoma.

Remapping significantly increased reading speed in simulated CFL subjects. Additional testing should examine the efficacy of remapping for reading and other visual tasks for patients with advanced CFL.

OBJECTIVE

The aim of this study was to compare long-term outcomes after anterior talofibular ligament (ATFL) and calcaneofibular ligament (CFL) repair and reconstruction at 5-10 years after surgery.

METHODS

Forty-five patients who underwent surgical repair or reconstruction of both ATFL and CFL were retrospectively investigated in this study. American Orthopedic Foot and Ankle Society (AOFAS), Karlsson Score, and Tegner activity scale were used to evaluate the ankle function at a follow-up of 5-10 years. Ultrasound examination was used to evaluate the ATFL and CFL, and MRI was used to evaluate the cartilage.

RESULTS

At final follow-up, no patient had recurrent ankle instability. There were no significant differences in AOFAS (92.6 ± 6.5 vs 89.6 ± 3.4; n.s.) or Karlsson Score (93 ± 8.2 vs 90.6 ± 5.0; n.s.) between the reconstruction group (twenty patients) and the repair group (twenty-five patients) postoperatively. There were also no significant differences in activity level as measured by the Tegner activity score (6 (range 4 to 8) vs 6 (range 5 to 7); n.s.). Five patients in the reconstruction group complained of some tightness of the ankles. Ultrasound showed the reconstructed ligaments maintained good continuity and were thicker than the repaired ligaments.

CONCLUSIONS

Patients in both the repair and the reconstruction cohort had high patient satisfaction with the outcomes and high function and activity levels that indicated recreational sports participation over a long period.

METHODS

III.

Herein, we developed an enantioselective addition of aliphatic aldehydes to 2-hydroxychalcone promoted by cooperative organocatalysts, giving access to hybrid flavonoids in excellent enantioselectivities. This reaction took advantage of cycloisomerization of 2-hydroxychalcone to form a transient flavylium under the irradiation of 24 W CFL, which was trapped by the in situ generated chiral enamine intermediate. The synergistic action of chiral phosphoric acid secured the excellent outcome of this reaction by ion-pairing with the transient flavylium.

The lattice Boltzmann method is a simulation technique in computational fluid dynamics. In its standard formulation, it is restricted to regular computation grids, second-order spatial accuracy, and a unity Courant-Friedrichs-Lewy (CFL) number. This paper advances the standard lattice Boltzmann method by introducing a semi-Lagrangian streaming step. The proposed method allows significantly larger time steps, unstructured grids, and higher-order accurate representations of the solution to be used. The appealing properties of the approach are demonstrated in simulations of a two-dimensional Taylor-Green vortex, doubly periodic shear layers, and a three-dimensional Taylor-Green vortex.

Background: Colorectal cancer (CRC) is among the deadliest cancers, wherein early dissemination of tumor cells, and consequently, metastasis formation, are the main causes of mortality and poor prognosis. Cofilin-1 (CFL-1) and its modulators, LIMK1/SSH1, play key roles in mediating the invasiveness by driving actin cytoskeleton reorganization in various cancer types. However, their clinical significance and prognostic value in CRC has not been fully explored. Here, we evaluated the clinical contribution of these actin regulators according to TNM and consensus molecular subtypes (CMSs) classification.

Methods: CFL-1, LIMK1 and SSH1 mRNA/protein levels were assessed by real-time PCR and immunohistochemical analyses using normal adjacent and tumor tissues obtained from a clinical cohort of CRC patients. The expression levels of these proteins were associated with clinicopathological features by using the chi square test. In addition, using RNA-Seq data of CRC patients from The Cancer Genome Atlas (TCGA) database, we determine how these actin regulators are expressed and distributed according to TNM and CMSs classification. Based on gene expression profiling, Kaplan-Meier survival analysis was used to evaluated overall survival.

Results: Bioinformatic analysis revealed that LIMK1 expression was upregulated in all tumor stages. Patients with high levels of LIMK1 demonstrated significantly lower overall survival rates and exhibited greater lymph node metastatic potential in a clinical cohort. In contrast, CFL-1 and SSH1 have expression downregulated in all tumor stages. However, immunohistochemical analyses showed that patients with high protein levels of CFL-1 and SSH1 exhibited greater lymph node metastatic potential and greater depth of local invasion. In addition, using the CMSs classification to evaluate different biological phenotypes of CRC, we observed that LIMK1 and SSH1 genes are upregulated in immune (CMS1) and mesenchymal (CMS4) subtypes. However, patients with high levels of LIMK1 also demonstrated significantly lower overall survival rates in canonical (CMS2), and metabolic (CMS3) subtypes.

Conclusions: We demonstrated that CFL-1 and its modulators, LIMK1/SSH1, are differentially expressed and associated with lymph node metastasis in CRC. Finally, this expression profile may be useful to predict patients with aggressive signatures, particularly, the immune and mesenchymal subtypes of CRC.

Keywords: Cofilin-1; Colorectal cancer; Consensus molecular subtypes; LIMK1; Lymph node metastasis; SSH1.

Since February-2020, the world has been battling a tragic public-health crisis with the emergence and spread of 2019-nCoV. Due to the lack of information about the pathogenesis-specific treatment of Covid-19, early diagnosis and timely treatment are important. However, there is still a lack of information about routine-blood-parameteres (RBP) findings and effects in the disease process. Although the literature includes various interventions, existing studies need to be generalized and their reliability improved. In this study, the efficacy of routine blood values used in the diagnosis and prognosis of Covid-19 and independent biomarkers obtained from them were evaluated retrospectively in a large patient group. Low lymphocyte (LYM) and white-blood-cell (WBC), high CRP and Ferritin were effective in the diagnosis of the disease. The (d-CWL) = CRPWBC∗LYM and (d-CFL) = CRP∗FerritinLYM biomarkers derived from them were the most important risk factors in diagnosing the disease and were more successful than direct RBP values. High d-CWL and d-CFL values largely confirmed the Covid-19 diagnosis. The most effective RBP in the prognosis of the disease was CRP. (d-CIT) = CRP*INR*Troponin; (d-CT) = CRP*Troponin; (d-PPT) = PT*Troponin*Procalcitonin biomarkers were found to be more successful than direct RBP values and biomarkers used in previous studies in the prognosis of the disease. In this study, biomarkers derived from RBP were found to be more successful in both diagnosis and prognosis of Covid-19 than previously used direct RBP and biomarkers.

Keywords: Biochemical, hematological and inflammatory biomarkers; Blood routine parameters; Covid-19; Covid-19′s ındependent biomarkers.

We present pressure-assisted capillary force lithography (CFL) to generate self-modulating polymer resist patterns without residual layers and film instability. The method utilizes roof collapse of a patterned, deformable poly(dimethyl siloxane) (PDMS) mold that is placed on a thermoplastic polymer film with a constant external pressure (approximately 4 bars) and the resulting shape-variable capillary filling of a polymer melt into the reduced void space. A constraint on the coated polymer layer thickness was derived in order to ensure that there is no residual layer left after patterning and at the same time that film stability is guaranteed without film dewetting within the cavity. In addition, the height of a polymer pattern at the center of the filled void was estimated as a function of initial polymer layer thickness based on the assumption of the hemispherical shape of a meniscus and full capillary rise, which agrees well with the experimental data.

Background: The 11-item Facial Line Outcomes (FLO-11) questionnaire is content validated for measuring the negative psychological impacts of crow's feet lines (CFL).

Objectives: The aims of this study were to determine psychological impacts of forehead lines (FHL) alone and upper facial lines (UFL: FHL + CFL + glabellar lines [GL]) and to assess adequacy of FLO-11 to measure these impacts.

Methods: Participants aged at least 18 years participated in concept elicitation and cognitive interviews to identify and define psychological impacts of UFL. They completed the FLO-11 questionnaire to assess its ability to measure psychological impacts of facial lines and its comprehensiveness in doing so.

Results: Forty interviews were completed by 29 participants. Twenty participants each provided interviews for FHL and for UFL. The most commonly reported psychological impacts for FHL and UFL, respectively, were feeling unattractive (85%, 80%), looking less attractive than desired (85%, 70%), feeling bothered (80%, 70%), feeling good/bad about appearance (80%, 70%), looking older than actual age (75%, 65%), and feeling stressed (70%, 70%). For FHL, 70% of participants also reported looking older than desired as a psychological impact. More than 50% of participants agreed that all 11 FLO-11 items measured a psychological impact for FHL. More than 50% reported that 9 of 11 items measured a psychological impact for UFL. The majority of participants (FHL, 65%; UFL, 60%) reported that the FLO-11 questionnaire is comprehensive in measuring psychological impacts of facial lines.

Conclusions: FHL and UFL have psychological impacts on patients, and FLO-11 is a content valid, comprehensive instrument for measuring them.

It is clinically challenging to distinguish between ankle and subtalar joints instability in vivo. Understanding the changes in load-displacement at the ankle and subtalar joints after ligament injuries may detect specific changes in joint characteristics that cannot be detected by investigating changes in range of motion alone. The effect of restricting joints end range of motion with ankle braces was already established, but little is known about the effect of an ankle brace on the flexibility of the injured ankle and subtalar joints. Therefore, the purposes of this study were to (1) understand how flexibility is affected at the ankle and subtalar joints after sectioning lateral and intrinsic ligaments during combined sagittal foot position and inversion and during internal rotation and (2) investigate the effect of a semi-rigid ankle brace on the ankle and subtalar joint flexibility. Kinematics and kinetics were collected from nine cadaver feet during inversion through the range of ankle flexion and during internal rotation. Motion was applied with and without a brace on an intact foot and after sequentially sectioning the calcaneofibular ligament (CFL) and the intrinsic ligaments. Segmental flexibility was defined as the slope of the angle-moment curve for each 1 Nm interval. Early flexibility significantly increased at the ankle and subtalar joint after CFL sectioning during inversion. The semi-rigid ankle brace significantly decreased early flexibility at the subtalar joint during inversion and internal rotation for all ligament conditions and at the ankle joint after all ligaments were cut.