Thyroid function and autoimmunity abnormalities are frequently pr

Thyroid function and autoimmunity abnormalities are frequently present in children with VKC. Children with VKC should be screened for thyroid function and evaluated for thyroid autoimmunity.”
“The widespread availability of authoritative guidance Pfizer Licensed Compound Library cell line on prescribing from a wide variety of international and national bodies calls into question the need for additional local formulary advice. This article

describes contemporary local formulary management in the United Kingdom and discusses the areas where local decision making remains valuable. Local formularies can fulfil important roles which justify their continued existence, including ensuring local ownership see more and acceptance of advice, rapid dissemination of information, responsiveness to local circumstances and service design, sensitivity to local

pricing arrangements and close professional links with commissioners, pharmacists and prescribers.”
“This paper is concerned with addressing how plant tissue mechanics is related to the micromechanics of cells. To this end, we propose a mesh-free particle method to simulate the mechanics of both individual plant cells (parenchyma) and cell aggregates in response to external stresses. The model considers two important features in the plant cell: (1) the cell protoplasm, the interior liquid phase inducing hydrodynamic phenomena, and (2) the cell wall material, a viscoelastic solid material that contains the protoplasm. In this particle framework, the cell fluid is modeled by smoothed particle hydrodynamics (SPH), a mesh-free method typically used to address problems selleck chemicals llc with gas and fluid dynamics. In the solid phase (cell wall) on the other hand, the particles are connected by pairwise interactions holding them together and preventing the fluid to penetrate the cell wall. The cell wall hydraulic conductivity (permeability) is built in as well through the SPH formulation. Although this model is also meant to be able to deal with dynamic and even

violent situations (leading to cell wall rupture or cell-cell debonding), we have concentrated on quasi-static conditions. The results of single-cell compression simulations show that the conclusions found by analytical models and experiments can be reproduced at least qualitatively. Relaxation tests revealed that plant cells have short relaxation times (1 mu s-10 mu s) compared to mammalian cells. Simulations performed on cell aggregates indicated an influence of the cellular organization to the tissue response, as was also observed in experiments done on tissues with a similar structure.”
“Nonenveloped viral capsids are metastable structures that undergo conformational changes during virus entry that lead to interactions of the capsid or capsid fragments with the cell membrane.

The rate of PTEN alterations in melanoma cell lines, primary mela

The rate of PTEN alterations in melanoma cell lines, primary melanoma, and metastatic melanoma is 27.6, 7.3, and 15.2%, respectively. Three mutations were found in both melanoma cell lines and biopsies. These mutations

are scattered throughout the gene, with the exception of exon 9. A mutational hot spot is found in exon 5, which encodes the phosphatase activity domain. Evidence is also presented to suggest that numerous homozygous deletions and missense variants exist in the PTEN transcript. Studying PTEN functions and implications of its mutations and other genes could provide insights KU57788 into the precise nature of PTEN function in melanoma and additional targets for new therapeutic approaches.”
“Our inability to distinguish between low-grade prostate cancers that pose no threat and those that can kill compels newly diagnosed early prostate cancer patients to make decisions that may negatively affect their lives needlessly for years afterward. To reliably stratify patients into different risk categories and apply appropriate treatment,

we need a better molecular understanding of prostate cancer progression. Androgen ablation therapy and 5-alpha reductase inhibitors reduce dihydrotestosterone levels and increase apoptosis. Because of the differing biological potentials of tumor cells, however, these treatments may, in some cases, worsen outcome by selecting for or inducing adaptation of stronger androgen receptor signaling pathways. Reduced dihydrotestosterone also may be associated with altered survival pathways. Complicating treatment

effects further, molecular adaptation may be accelerated by interactions between BLZ945 epithelial and stromal cells. The hypothesis that early prostate cancer cells with differing biological potential may respond differently to finasteride treatment is worth testing. Ongoing studies using a systems biology approach in a preoperative prostate cancer setting are testing this hypothesis toward developing more-rational clinical interventions.”
“SETTING: The Korean Institute of Tuberculosis, Seoul, Republic of Korea.\n\nOBJECTIVE: To develop a simple, Selleckchem P505-15 direct drug susceptibility testing (DST) technique using Kudoh-modified Ogawa (KMO) medium.\n\nDESIGN: The critical concentrations of isoniazid (INH), rifampicin (RMP), kanamycin (KM) and ofloxacin (OFX) for KMO medium were calibrated by comparing the minimal inhibitory concentrations (MICs) against clinical isolates of Mycobacterium tuberculosis on KMO with those on Lowenstein-Jensen (q). The performance of the direct KMO DST technique was evaluated on 186 smear-positive sputum specimens and compared with indirect LJ DST.\n\nRESULTS: Agreement of MICs on direct vs. indirect DST was high for INH, RMP and OFX. KM MICs on KMO were 10 mu g/ml higher than those on LJ. The critical concentrations of INH, RAP, OFX and KM for KMO were therefore set at 0.2, 40.0, 2.0, and 40.0 mu g/ml.

) Hale and P clavuliferum (Rasanen) Streimann The data showed th

) Hale and P clavuliferum (Rasanen) Streimann. The data showed that the species are anatomically similar, including the presence of epicortex, the upper cortex anatomy and the characteristics of rhizines and ciliae. In the medulla of the two species there are star-shaped clusters of hyphae associated with

the presence of salazinic acid. This study showed that the anatomical characteristics are constant for the Parmotrema group studied.”
“Objectives To examine the relationships between plasma and tissue markers of systemic and vascular inflammation and obesity and insulin resistance and determine the effects of aerobic exercise training plus weight loss (AEX+WL) and weight loss (WL) alone on these biomarkers. Design Prospective controlled study. Setting Veterans Affairs NVP-AUY922 Medical Center and University research setting. Participants Overweight and obese sedentary postmenopausal women (N=77). Interventions Six months, 3d/wk AEX+WL (n= 37) or WL (n=40). Measurements Total-body dual-energy X-ray absorptiometry, abdominal computed tomography, hyperinsulinemic-euglycemic clamps (a criterion standard method of assessing insulin sensitivity), adipose tissue biopsies (n=28), and blood for homeostasis model assessment-insulin resistance, and soluble forms of intracellular adhesion molecule

1 (sICAM-1) and vascular cell adhesion molecule 1 Selleck HSP990 (sVCAM-1), C-reactive protein (CRP), and serum amyloid A (SAA). Results Body weight SBC-115076 mw (P smaller than .001), percentage of fat (P smaller than .001), visceral fat (P smaller than .005), triglyceride levels (P smaller than .001), and systolic blood pressure decreased comparably after WL and AEX+WL (P=.04). Maximal oxygen consumption increased 16% after AEX+WL (P smaller than .001). Insulin resistance decreased in both groups (P=.005). Glucose utilization according to the clamp increased 10% (P=.04) with AEX+WL and 8% with WL (P=.07). AEX+WL decreased CRP by 29% (P smaller than .001) and WL by 21% (P=.02). SAA levels decreased twice as much after AEX+WL (-19%, P=.02) as after WL

(-9%, P=.08). Plasma sICAM-1 and sVCAM-1 levels did not change, but women with the greatest reduction in plasma sICAM-1 levels had the greatest reductions in fasting glucose (P=.02), insulin (P=.02), and insulin resistance (P=.004). Gluteal ICAM messenger ribonucleic acid levels decreased 27% after AEX+WL (P=.02) and did not change after WL. Conclusion Obesity and insulin resistance worsen markers of systemic and vascular inflammation. A reduction in plasma sICAM-1 is important to improve insulin sensitivity. CRP, SAA, and tissue ICAM decrease with exercise and weight loss, suggesting that exercise training is a necessary component of lifestyle modification in obese postmenopausal women.”
“Micro-organisms react to a rapid temperature downshift by triggering a physiological response to ensure survival in unfavourable conditions.

In order to deal with this challenge in planning and control of l

In order to deal with this challenge in planning and control of logistics activities, several solutions have been introduced. One of the most latest one

is the application of autonomy. The paradigm of autonomy in inbound logistics, can be reflected in decisions for real-time scheduling and control of material flows. Integration of autonomous control with material carrier objects can realize the expected advantages of this alternative into shop-floors. Since pallets Blebbistatin research buy (bins, fixtures, etc.) are some common used carrier objects in logistics, they have the potential to undertake the responsibility of real-time jobs dispatching to machines in the shop-floor scheduling problem. Hence, the current paper covers the problem of real-time scheduling in a stochastic and complex shop-floor environment, by means of autonomy. Indeed, the sustainment’s advantage of pallets in manufacturing systems has inspired the idea of developing learning pallets (Lpallets) with the capability of autonomous GS-1101 supplier control in complex and uncertain

logistics environment with abrupt changes. Among some intelligent techniques, the artificial neural network (ANN) and, specially, the radial basis function network (RBFN) is selected to transmit the abilities of intelligent decision-making as well as learning to Lpallets in a distributed manner. Some variants in training and RBFN application alternatives are considered to evaluate the competency of RBFN for Lpallets. An Lpallet makes its dispatching and control decision based on its own experience and intelligence about the entire system and local situations in an

exemplary hybrid flow-open shop problem. To prove the claimed application of RBFN in autonomous Lpallets a discrete-event 3-deazaneplanocin A mw simulation model is developed for the assembly scenario. (C) 2012 Elsevier B.V. All rights reserved.”
“Although the injury risks of boxing is well known, this sport continuous to attract athletes and an increase of introduction to boxing is observed in the last decade. In terms of injury locations, head and face are reported as most common sports. Present study aimed to examine the hearing differences of elite amateur boxers as a result of punch strokes in boxing.\n\nSubjects are interested in active boxing for 5-14 years (mean 6.67) and between the age of 18-32 (mean 21.71). Screened group consisted of 21 male boxers. Auditory brainstem responses, pure tone and high frequency audiogram tests were conducted for boxers and unscreened groups in the standard acoustically controlled rooms using Interacoustics Clinical Computer Audiometer.\n\nMean +/- standard deviations are reported. Groups were compared by Student’s t test p<0.05. Auditory brainstem responses and pure tone values were determined in range of I-V inter-pick latency (ms).

Radiation exposure was 7 times lower during the suspension test

Radiation exposure was 7 times lower during the suspension test. Cobb angle reductions achieved in the proximal and main curves by the two tests were similar. The traction test achieved greater reduction of the distal curve. Flexibility in the axial plane and in the sagittal plane was analyzed with the suspension test. The reduction in apical rotation was not correlated with the reduction in Cobb angle. The EOS suspension test can be used for the assessment of spinal flexibility in AIS. It gives a global

vision of the deformity with new flexibility indices in the axial and sagittal planes.”
“Elk-1 is a member of the E-twenty-six (ETS) domain superfamily of transcription factors and has been traditionally associated with mitogen-induced immediate early gene transcription upon phosphorylation by mitogen activated protein kinases (ERK/MAPK). Elk-1 is not only upregulated but also phosphorylated Selleckchem MI-503 in brain tumour cells. However,

in this study, we S3I-201 show for the first time that S383-phosphorylated Elk-1 (P-S383-Elk-1) is associated with mitotic spindle poles from metaphase through telophase and relocates to the spindle midbody during cytokinesis, while Thr417Ala mutation is associated with DNA throughout mitosis. Serine 383 phosphorylation appears to be important for polar localization of Elk-1, since exogenous protein including serine-to-alanine mutation was seen to be distributed throughout the spindle fibres. We further show that Elk-1 interacts with the cell cycle kinase Aurora-A, and when Aurora inhibitors are used, P-S383-Elk-1 fails to localize to the poles and remains associated with DNA. Apart from one transcriptional repressor molecule, Kaiso, this is the first time a transactivator was shown to possess such mitotic localization and interaction. The functional significance and detailed mechanism of this

cell cycle-related localization of Elk-1 are yet to be determined. Copyright (c) 2013 John Wiley & Sons, Ltd.”
“Background Hemiplegia and hemiparesis are the most common deficits caused by stroke. A few small clinical trials suggest that fluoxetine enhances motor recovery but its clinical efficacy is unknown. We therefore aimed to investigate whether fluoxetine would enhance motor recovery if given soon after an ischaemic stroke to patients who A-1210477 solubility dmso have motor deficits.\n\nMethods In this double-blind, placebo-controlled trial, patients from nine stroke centres in France who had ischaemic stroke and hemiplegia or hemiparesis, had Fugl-Meyer motor scale (FMMS) scores of 55 or less, and were aged between 18 years and 85 years were eligible for inclusion. Patients were randomly assigned, using a computer random-number generator, in a 1:1 ratio to fluoxetine (20 mg once per day, orally) or placebo for 3 months starting 5-10 days after the onset of stroke. All patients had physiotherapy. The primary outcome measure was the change on the FMMS between day 0 and day 90 after the start of the study drug.

In addition, suppression of miR-27a in HBERST cells attenuated th

In addition, suppression of miR-27a in HBERST cells attenuated the capacity of such cells to grow in an anchorage-independent manner. We also found that suppression of the PP2A B56 gamma expression resulted in upregulation of miR-27a similar to that achieved by the introduction of ST, indicating that dysregulation of miR-27a expression in ST-expressing cells was mediated by the ST-PP2A interaction. Moreover, we discovered that Fbxw7 gene encoding F-box/WD repeat-containing protein 7 was a potential miR-27a target validated by dual-luciferase reporter system analysis. The inverse correlation between

miR-27a expression levels and Fbxw7 protein expression was further confirmed in both cell models Batimastat mouse and human tumor samples. Fbxw7 regulates cell-cycle progression through the ubiquitin-dependent proteolysis of a set H 89 research buy of substrates, including c-Myc, c-Jun, cyclin E1 and Notch 1. Thus, promotion of cell growth arising from the suppression of Fbxw7 by miR-27a overexpression might be responsible for the viral oncoprotein ST-induced malignant transformation. These observations demonstrate that miR-27a functions as an oncogene in human tumorigenesis. Oncogene (2011) 30, 3875-3886; doi:10.1038/onc.2011.103; published online 4 April 2011″
“The structure of modern clinical trials is designed to protect patient safety while generating safety and efficacy

data. Safety is the primary concern, and United States regulations are shaped by a series of responses to incidents, including notable safety lapses and unethical trials.

These regulations focus on 3 essential components, AC220 solubility dmso defined by the 1979 Belmont Report: respect for persons, beneficence, and justice. Further, the international community has formally outlined good clinical practice (GCP), which mandates that trials are designed to produce meaningful data, conform to international ethics regulations, and provide assurances that data are reported in a credible and reliable manner. The Food and Drug Administration (FDA) and federal government have outlined the necessary components of clinical trials in the Code of Federal Regulations (CFR). These include institutional review boards (IRBs), standard operating procedures (SOPs), sites, sponsors, investigators, and patients. The investigator is the center of the trial and is required to sign an agreement with the federal government to uphold the CFR. Investigator duties include making sure that investigator and support staff having appropriate qualifications, delegating duties, monitoring the study for compliance and record keeping, providing care, and accepting accountability for the trial, among other duties. Physicians, who already have significant time demands, need a well-trained staff, including clinical coordinators, to adequately meet these duties.

The effects of the KA on the sleep time were observed using a hyp

The effects of the KA on the sleep time were observed using a hypnosis test, and the tail-withdrawal latency was analyzed using the tail-withdrawal test. In the hypnosis test, KA (2.5, 5 or 10 ng; icy administered) treatment had no distinctive effects on the sleep time of mice treated with emulsified inhalation anesthetics. In the tail-withdrawal test, KA (0.2, 0.4 or 0.8 ng; it administered) treatment significantly and dose-dependently decreased the tail-withdrawal

latency of mice treated with emulsified anesthetics. These results suggested that KA receptors may modulate the analgesic but not hypnotic effects induced by emulsified en flurane, isoflurane or sevoflurane.”
“Lesch-Nyhan disease (LND), a genetic disorder associated with motor and psychiatric disturbance and self-injurious behaviour (SIB) is Caused by a complete deficiency of hypoxanthine-guanine compound screening assay phosphoribosyltransferase

(HPRT). The connection between enzyme deficiency and neurological involvement is still unclear. Evidence exists for a role of basal ganglia dysfunction with decreased dopamine and excess serotonin selleck screening library striatal content. In this Study, we investigate the role of serotonin receptor 2C (HTR2C) in the brains of HPRT gene knock-out mice, a model of LND. HTR2C expression is analyzed by real-time polymerase chain reaction (PCR) using SYBR-green detection methods. The percentage of edited HTR2C mRNA was determined by direct sequencing of amplification products of the region containing the editing sites. We found a 55% increase in the expression of HTR2C gene but no significant difference in mRNA editing levels between knock-out and control mice. The above alteration found in HPRT-deficient mice is similar to

those found in other animal models used to Study aggressive and self injurious behaviour. (C) 2009 Elsevier Ltd. All rights reserved.”
“Premature luteal demise or luteal insufficiency is not well characterised LBH589 as a cause of pregnancy loss in domestic species, including horses. In this report, a mare inseminated with cooled-transported semen at our facility returned for a routine pregnancy diagnosis at 15 days post ovulation. Ultrasonography per rectum revealed endometrial oedema and the absence of visual indication of a corpus luteum on either ovary. Nonetheless, an embryonic vesicle small for the gestational age was identified. Daily oral altrenogest treatment was implemented immediately. Serum progesterone concentration was 0.67 ng/ml, which is below the threshold considered adequate for pregnancy maintenance in the mare. Examinations were repeated at 17, 25, 30, 39, 49, 72 and 120 days post ovulation. At 25 days post ovulation the embryonic vesicle presented normal development for the gestational age. In addition, sequential blood samples were collected to measure progesterone, equine chorionic gonadotrophin and oestrone sulphate concentrations.


“Background: Pseudomonas aeruginosa is a common bacterium


“Background: Pseudomonas aeruginosa is a common bacterium that can cause disease in humans and other animals. This study was conducted to screen for molecular detection and antimicrobial-resistant P. aeruginosa in Musca domestica in different locations in the Iranian provinces of Shahrekord and Isfahan. Methods: Musca domestica were captured by both manual and sticky trap methods,

during the daytime, from household kitchens, cattle farms, animal hospitals, human hospitals, slaughterhouses and chicken farms at random locations in Shahrekord and Isfahan provinces of Iran, and subsequently transported to the laboratory for detection of P. aeruginosa. In the laboratory, flies were identified and Dinaciclib chemical structure killed by refrigeration in a cold chamber at -20 degrees C, then placed in 5 mL peptone water and left at room temperature for five hours before being processed. Anlotinib Pseudomonas isolates were preliminarily identified to genus level based on colony morphology and gram staining, and their identity was further

confirmed by polymerase chain reaction. Results: Overall blaTEM gene was recovered from 8.8 % (53/600) of the P. aeruginosa isolated from houseflies collected from the two provinces. A slightly higher prevalence (10.7 %; 32/300) was recorded in Shahrekord province than Isfahan province (7.0 %; 21/300). The locations did not differ statistically (p smaller than 0.05) in bacterial prevalence in flies. Seasonal prevalence showed a significantly lower infection frequency during autumn. Conclusions: Houseflies are important in the epidemiology of P. aeruginosa infections.”
“Traumatic brain injury

(TBI) causes substantial morbidity and mortality worldwide. A key component of both mild and severe TBI is diffuse axonal injury. Except in cases of extreme mechanical strain, when axons are torn at the moment of trauma, axonal stretch injury is characterized by early cytoskeletal proteolysis, transport disruption, and secondary axotomy. Calpains, a family of Ca2+-dependent proteases, have been implicated in this pathologic cascade, but direct in vivo evidence is lacking. To test the hypothesis that calpains play a causal role in axonal stretch injury in vivo, we used selleck our rat optic nerve stretch model following adeno-associated viral (AAV) vector-mediated overexpression of the endogenous calpain inhibitor calpastatin in optic nerve axons. AAV vectors were designed for optimal expression of human calpastatin (hCAST) in retinal ganglion cells (RGCs). Calpain inhibition by the expressed protein was then confirmed in primary cortical cultures. Finally, we performed bilateral intravitreal injections of AAV vectors expressing hCAST or the reporter protein ZsGreen 3 weeks prior to unilateral optic nerve stretch.

Interestingly, most of this progress has been made by understandi

Interestingly, most of this progress has been made by understanding DNA that does not encode genes.”
“This article describes technological advances in quadrature transverse electromagnetic (TEM) volume coils and phased arrays reported recently from our laboratory developed for MRI and MRS imaging of the human brain. The first part of this work presents a new method for tuning TEM volume coils based https://www.selleckchem.com/products/z-devd-fmk.html on measurements of the radiofrequency current distribution in the coil elements. This technique facilitates bench adjustment of the coils’ homogeneity and is particularly important for tuning double-tuned TEM volume coils.

We have also used this method to https://www.selleckchem.com/products/smoothened-agonist-sag-hcl.html optimize other TEM configurations such as a quadrature TEM half-volume coil and a split TEM coil. TEM half-volume coils provide greater sensitivity over localized regions than conventional full-volume coils, and the split TEM coil provides greater patient access and ease of use. The second part of this work

describes the development of single-tuned and double-tuned transmit TEM volume coils in combination with phased arrays. A variety of different techniques for active detuning of single-tuned and double-tuned TEM volume coils are presented along with the development of phased arrays and transmission line preamplifier decoupling. The final section describes the use of counter rotating current (CRC) surface coils in phased arrays. Because of the intrinsic isolation of CRC Selleck A 1155463 coils from transmit volume coils, CRC arrays can be used simultaneously with volume coils

for both reception and transmission. Near the center of the human head where both the phased array and the volume coil produce similar sensitivities, simultaneous reception enhances the signal-to-noise ratio. Conversely, simultaneous transmission can be used to boost the transmit field in peripheral brain regions from the volume coil to provide a more homogeneous transmit field. Copyright (C) 2008 John Wiley & Sons, Ltd.”
“Objective: To evaluate the efficacy of pramipexole on polysomnographic measures, patient ratings and a clinical rating in Japanese patients with primary restless legs syndrome (RLS).\n\nMethods: Patients with moderate to severe RLS having periodic limb movements in bed index (PLMI) >= 5 were randomly assigned to receive pramipexole or placebo in a 6-week, double-blind, placebo-controlled study with forced titration from 0.125 to 0.75 mg/day. Both polysomnography (PSG) and the suggested immobilization test (SIT) were performed at baseline and 6 weeks after starting treatment.\n\nResults: The analysis of covariance of log-transformed PLMI showed that the adjusted means at the end of study were significantly smaller in the pramipexole group than in the placebo group (p = 0.0019).

A growing body of data indicates that Claudin expression is alter

A growing body of data indicates that Claudin expression is altered in numerous epithelial cancers in a stage- and LY2606368 concentration tumor-specific manner. While a single universal mechanism is still lacking, accumulating evidence supports a role for epigenetic regulation of Claudin expression in tumorgenesis, with concomitant alterations in barrier function. We review here new insights and challenges in understanding Claudin

function in normal physiology and cancer. (C) 2011 Elsevier B.V. All rights reserved.”
“DNA reprogramming by DNA (cytosine-5)-methyltransferases (dnmts) after fertilisation is a dynamic mechanism that is essential for early development. Amongst the three types of dnmt genes in vertebrates, dnmt3 is the one involved in de novo methylation and comprises three related genes, termed dnmt3a, dnmt3b and dnmt3L in mammals. Zebrafish (Danio redo) has six dnmt3 paralogues, which have hitherto

been termed dnmt3 to dnmt8. Bayesian inference of phylogeny and synteny GW4869 analysis revealed that dnmt6 and dnmt8 are in fact duplicated dnmt3a genes, whereas the other paralogues are closely related to dnmt3b. Hence, we propose a revised nomenclature that more accurately reflects the relationship amongst zebrafish dnmt3 genes. Both dnmt3a genes were ubiquitously expressed in adult tissues, whilst the various dnmt3b paralogues were differentially

expressed, with notably high expression levels in the gonads. The influence of embryonic temperature on dnmt3 expression was investigated, since it is known to have a significant impact in early development and a long-term effect on growth in some teleost species. Embryos were incubated at 23, 27 or 31 degrees C and samples collected at six developmental stages from blastula until protruding mouth. Dnmt3 expression during early development was remarkably dynamic. In particular, mRNA levels of the two dnmt3a genes showed a marked increase throughout development and several significant differences in dnmt3a and dnmt3b transcript levels were found between temperatures at the same developmental point. Taken together, our data indicate that dnmt3 paralogues Caspase activity assay are diverging and that dnmt3a and dnmt3b may play different roles in thermal epigenetic regulation of gene expression during early development. (c) 2012 Elsevier B.V. All rights reserved.”
“High quality gamete production in males and females requires the pituitary gonadotropin follicle stimulating hormone (FSH). In this report a novel chemical class of small molecule inhibitors of FSH receptor (FSHR) is described. ADX61623, a negative allosteric modulator (NAM), increased the affinity of interaction between (125)I-hFSH and human FSHR (hFSHR) five fold.