Office-Based Rapid Diagnosis and Management of External Ophthalmomyiasis Via Smartphone-Aided Innovative Tools in an Under-Resourced Setting.

PURPOSE: To evaluate the use of smartphone-based innovative tools named Anterior segment photography with intraocular lens (ASPI) and smartphone based intraocular lens microscope (IOLSCOPE) to demonstrate, treat, and diagnose the patients of ophthalmomyiasis caused by Oestrus ovis. METHODS: A retrospective interventional case series of six patients infested with O. ovis presenting in a rural health center with symptoms of burning, itching, watering, and foreign body sensation diagnosed with ophthalmomyiasis. Diagnosis and treatment were performed using an innovative imaging technique that is, an optical system formed by intraocular lens (IOL) and smartphone. RESULTS: The smartphone-based ASPI clearly detected and removed the moving larvae from the ocular surface, followed by microbiological diagnosis of O.ovis larvae with the help of an IOLSCOPE. CONCLUSION: The importance of ASPI and IOLSCOPE for the management of ophthalmomyiasis in peripheral health centers devoid of slitlamp and microscopes has been emphasized here.

Covalent functionalization of polypropylene filters with diazirine-photosensitizer conjugates producing visible light driven virus inactivating materials.

The SARS-CoV-2 pandemic has highlighted the weaknesses of relying on single-use mask and respirator personal protective equipment (PPE) and the global supply chain that supports this market. There have been no major innovations in filter technology for PPE in the past two decades. Non-woven textiles used for filtering PPE are single-use products in the healthcare environment; use and protection is focused on preventing infection from airborne or aerosolized pathogens such as Influenza A virus or SARS-CoV-2. Recently, C-H bond activation under mild and controllable conditions was reported for crosslinking commodity aliphatic polymers such as polyethylene and polypropylene. Significantly, these are the same types of polymers used in PPE filtration systems. In this report, we take advantage of this C-H insertion method to covalently attach a photosensitizing zinc-porphyrin to the surface of a melt-blow non-woven textile filter material. With the photosensitizer covalently attached to the surface of the textile, illumination with visible light was expected to produce oxidizing 1O2/ROS at the surface of the material that would result in pathogen inactivation. The filter was tested for its ability to inactivate Influenza A virus, an enveloped RNA virus similar to SARS-CoV-2, over a period of four hours with illumination of high intensity visible light. The photosensitizer-functionalized polypropylene filter inactivated our model virus by 99.99% in comparison to a control.

A preliminary investigation into the design of pressure cushions and their potential applications for forearm robotic orthoses.

BACKGROUND: Load cells are often used in rehabilitation robotics to monitor human-robot interaction. While load cells are accurate and suitable for the stationary end-point robots used in rehabilitation hospitals, their cost and inability to conform to the shape of the body hinder their application in developing affordable and wearable robotic orthoses for assisting individuals in the activities of daily living. This exploratory work investigates the possibility of using an alternative technology, namely compliant polymeric air cushions, to measure interaction forces between the user and a wearable rigid structure. METHODS: A polymeric air cushion was designed, analyzed using a finite element model (FEM), and tested using a bench-top characterization system. The cushions underwent repeatability testing, and signal delay testing from a step response while increasing the length of the cushion's tubes. Subsequently, a 3D printed wrist brace prototype was integrated with six polymeric air cushions and tested in static conditions where a volunteer exerted isometric pronation/supination torque and forces in vertical and horizontal directions. The load measured by integrating data recorded by the six sensors was compared with force data measured by a high quality load cell and torque sensor. RESULTS: The FEM and experimental data comparison was within the error bounds of the external differential pressure sensor used to monitor the pressure inside the cushion. The ratio obtained experimentally between the pressure inside the pressure cushion and the 8 N applied load deviated by only 1.28% from the FEM. A drift smaller than 1% was observed over 10 cycles. The rise times of the cushion under an 8 N step response for a 0.46, 1.03, and 2.02 m length tube was 0.45, 0.39, and 0.37 s. Tests with the wrist brace showed a moderate root mean square error (RMSE) between the force estimated by the pressure cushions and the external load cells. Specifically, the RMSE was 13 mNm, 500 mN, and 1.24 N for forearm pronation/supination torque, vertical force, and horizontal force, respectively. CONCLUSIONS: The use of compliant pressure cushions was shown to be promising for monitoring interaction forces between the forearm and a rigid brace. This work lays the foundation for the future design of an array of pressure cushions for robotic orthoses. Future research should also investigate the compatibility of these polymeric cushions for data acquisition during functional magnetic resonance imaging in shielded rooms.

Bimanual elbow exoskeleton: Force based protocol and rehabilitation quantification.

An aging population, along with the increase in cardiovascular disease incidence that accompanies this demographic shift, is likely to increase both the economic and medical burden associated with stroke in western societies. Rehabilitation, the standard treatment for stroke, can be expanded and augmented with state of the art technologies, such as robotic therapy. This paper expands upon a recent work involving a force-feedback master-slave bimanual exoskeleton for elbow rehabilitation, named a Bimanual Wearable Robotic Device (BWRD). Elbow force data acquired during the execution of custom tasks is analyzed to demonstrate the feasibility of tracking patient progress. Two training tasks that focus on applied forces are examined. The first is called "slave arm follow", which uses the absolute angular impulse as a metric; the second is called "conditional arm static", which uses the rise time to target as a metric, both presented here. The outcomes of these metrics are observed over three days.

Moving toward automatic and standalone delineation of seismocardiogram signal.

The purpose of this research is to propose an algorithm that could accomplish automatic delineation of the seismocardiogram (SCG) signal without using a reference electrocardiogram R-wave. As a result, the SCG signal could be used, as a stand-alone solution for many cardiovascular medical applications such as hemorrhage detection, cardiac computed tomographic gating, cardiac resynchronization therapy, hemodynamics estimations and diastolic timed vibration. Multiple envelopes were derived from the seismocardiogram signal by using filtering and triple integration. The first envelope is referred as the heart rate envelope, which has the characteristics of having a period of exactly one cardiac cycle and its purpose is to replace the ECG R-wave as a reference point. Our dataset is based on the lower body negative pressure (LBNP) test that was conducted on 18 individuals, containing 21610 cardiac cycles. For 94% of the LBNP dataset, the aforementioned envelope estimated heart rate within 3 beats per minute. Three different peaks of the SCG signal are of our interest: isovolumic contraction (IM), aortic valve opening (AO) and aortic valve closure (AC). For each of these desired peaks of the SCG signal, a different envelope was designed in a manner that its peak is very close to IM, AO and AC, respectively. For the same lower body negative pressure data set, a mean difference of (9, 9, 6) and standard deviation of (8, 9, 9) millisecond between the peak of envelopes and IM, AO and AC is accomplished. This could be used as a good initial estimation of the annotation points.

Material versatility using replica molding for large-scale fabrication of high aspect-ratio, high density arrays of nano-pillars.

Arrays of high aspect-ratio (AR) nano-pillars have attracted a lot of interest for various applications, such as for use in solar cells, surface acoustic sensors, tissue engineering, bio-inspired adhesives and anti-reflective surfaces. Each application may require a different structural material, which can vary in the required chemical composition and mechanical properties. In this paper, a low cost fabrication procedure is proposed for large scale, high AR and high density arrays of nano-pillars. The proposed method enables the replication of a master with high fidelity, using the subsequent replica molds multiple times, and preparing arrays of nano-pillars in a variety of different materials. As an example applied to bio-inspired dry adhesion, polymeric arrays of nano-pillars are prepared in this work. Thermoset and thermoplastic nano-pillar arrays are examined using an atomic force microscope to assess their adhesion strength and its uniformity. Results indicate the proposed method is robust and can be used to reliably prepare nano-structures with a high AR.

A new seismocardiography segmentation algorithm for diastolic timed vibrations.

An algorithm based on the combination of electrocardiography (ECG) and seismocardiogranphy (SCG) is used to detect the start and the end of diastole in diastolic timed vibrations (DTV). The proposed algorithm uses the ECG-R wave as the reference point and detects the aortic valve closure (AC) and mitral valve closure (MC) points of the SCG signal. This algorithm enables DTV to operate very efficiently in comparison with previous ECG based algorithm. Prediction rate of 95 and 88 percent was achieved for detection of SCG-MC and SCG-AC respectively.

Precordial acceleration signals improve the performance of diastolic timed vibrations.

BACKGROUND AND OBJECTIVE: This paper introduces a seismocardiography based methodology of predicting the start and the end of diastole to be used in diastolic timed vibrations (DTV), which provides non-invasive emergency treatment of acute coronary thrombosis by applying direct mechanical vibrations to the patient chest during diastole of heart cycles. It is proposed that seismocardiogram (SCG), in combination with electrocardiogram (ECG), provides a new means of diastole prediction. METHODS: An accelerometer was placed on the sternum of 120 healthy participants and 22 ischemic heart patients to record precordial accelerations created by the heart. The accelerometer signal was used to extract SCG and phonocardiogram (PCG). Two independent trained experts annotated the extracted signals based on the timings of the start and end of diastole. RESULTS: In the ischemic heart disease population by using 15 consecutive SCG cycles, the start and end of diastole was predicted in the upcoming cycles with 95 percentile error margin of 10.7 ms and 5.8 ms, respectively. These error margins were 7.4 ms and 3.5 ms, respectively, for normal participants. CONCLUSION: The results provide that prediction of the aortic valve closure point in the SCG signal helps start the vibrator in time to cover most of the isovolumic relaxation period. Also, through prediction of the mitral valve closure point in the SCG signal, safety of the technique can be assessed through prediction of the amount of unwanted vibrations applied during the isovolumic contraction period.

Preferred patterns of diastolic timed vibrations for pre-hospitalization treatment of acute coronary ischemia.

This paper presents and evaluates preferred patterns of vibrations and active breaking techniques for the Diastolic Timed Vibrator (DTV). DTV uses low frequency mechanical vibrations applied to the chest to help in clot dissolution in pre-hospitalization treatment of acute coronary ischemia. In this work, we argue that random and ramp type vibration patterns increase the performance of the DTV method. Furthermore, we present results for various methods of vibration stopping aiming at reduction of vibration overspill into the systole of heart cycle of the patient.

Elastic anisotropy and low-temperature thermal expansion in the shape memory alloy Cu-Al-Zn.

Cu-based shape memory alloys are known for their technologically important pseudo-elastic and shapememory properties, which are intimately associated with the martensitic transformation. A combination of deformation theory and finite-strain elasticity theory has been employed to arrive at the expressions for higher order elastic constants of Cu-Al-Zn based on Keating's approach. The second- and third-order elastic constants are in good agreement with the measurements. The aggregate elastic properties like bulk modulus, pressure derivatives, mode Grüneisen parameters of the elastic waves, low temperature limit of thermal expansion, and the Anderson-Grüneisen parameter are also presented.

Simultaneous quantitation of rosuvastatin and gemfibrozil in human plasma by high-performance liquid chromatography and its application to a pharmacokinetic study.

A simple, sensitive and specific high-performance liquid chromatography method is described for simultaneous determination of rosuvastatin (RST) and gemfibrozil (GFZ) in human plasma using celecoxib as an internal standard (IS). The assay procedure involved extraction of RST, GFZ and IS from plasma into acetonitrile. Following separation and evaporation of the organic layer the residue was reconstituted in the mobile phase and injected onto an X-Terra C(18) column (4.6 x 150 mm, 5.0 microm). The chromatographic run time was less than 20 min using flow gradient (0.0-1.60 mL/min) with a mobile phase consisting of 0.01 M ammonium acetate:acetonitrile:methanol (50:40:10, v/v/v) and UV detection at 275 nm. Nominal retention times of RST, GFZ and IS were 6.7, 13.9 and 16.4 min, respectively. Absolute recovery of both analytes and IS was greater than 90%. The lower limit of quantification (LLOQ) of RST and GFZ was 0.03 and 0.30 microg/mL, respectively. Linearity was excellent (r(2) = 0.999) in the 0.03-10 microg/mL and 0.3-100 microg/mL ranges for RST and GFZ, respectively. The inter- and intra-day precisions in the measurement of RST quality control (QC) samples 0.03, 0.09, 2.50 and 8.00 microg/mL were in the range 2.37-9.78% relative standard deviation (RSD) and 0.92-10.08% RSD, respectively. Similarly, the inter- and intra-day precisions in the measurement of GFZ quality control (QC) samples 0.30, 0.90, 25.0 and 80.0 microg/mL were in the ranges 2.79-6.27 and 0.96-9.69% RSD, respectively. Accuracies in the measurement of QC samples for RST and GFZ were in the range 85.43-107.23 and 84.98-102.35% respectively, of the nominal values. RST and GFZ were stable in the array of stability studies viz., bench-top, auto-sampler and freeze-thaw cycles. Stability of RST and GFZ was established for 1 month at -80C. The application of the assay in an oral pharmacokinetic study in rats co-administered with RST and GFZ is described.

Determination of rosuvastatin in rat plasma by HPLC: Validation and its application to pharmacokinetic studies.

A specific, accurate, precise and reproducible high-performance liquid chromatography (HPLC) method was developed for the estimation of rosuvastatin (RST), a novel, synthetic and potent HMG-CoA inhibitor in rat plasma. The assay procedure involved simple liquid-liquid extraction of RST and internal standard (IS, ketoprofen) from a small plasma volume directly into acetonitrile. The organic layer was separated and evaporated under a gentle stream of nitrogen at 40 degrees C. The residue was reconstituted in the mobile phase and injected onto a Kromasil KR 100-5C18 column (4.6 x 250 mm, 5 microm). Mobile phase consisting of 0.05 m formic acid and acetonitrile (55:45, v/v) was used at a flow rate of 1.0 mL/min for the effective separation of RST and IS. The detection of the analyte peak was achieved by monitoring the eluate using a UV detector set at 240 nm. The ratio of peak area of analyte to IS was used for quantification of plasma samples. Nominal retention times of RST and IS were 8.6 and 12.5 min, respectively. The standard curve for RST was linear (r2 > 0.999) in the concentration range 0.02-10 microg/mL. Absolute recoveries of RST and IS were 85-110 and >100%, respectively, from rat plasma. The lower limit of quantification (LLOQ) of RST was 0.02 microg/mL. The inter- and intra-day precisions in the measurement of quality control (QC) samples, 0.02, 0.06, 1.6 and 8.0 microg/mL, were in the range 7.24-12.43% relative standard deviation (RSD) and 2.28-10.23% RSD, respectively. Accuracy in the measurement of QC samples was in the range 93.05-112.17% of the spiked nominal values. Both analyte and IS were stable in the battery of stability studies, viz. benchtop, autosampler and freeze-thaw cycles. RST was found to be stable for a period of 30 days on storage at -80 degrees C. The application of the assay to determine the pharmacokinetic disposition after a single oral dose to rats is described.

Simultaneous quantitation of etoricoxib, salicylic acid, valdecoxib, ketoprofen, nimesulide and celecoxib in plasma by high-performance liquid chromatography with UV detection.

A specific, accurate, precise and reproducible high performance liquid chromatography (HPLC) method was developed and validated for the simultaneous quantitation of etoricoxib, salicylic acid, valdecoxib, ketoprofen, nimesulide and celecoxib in human plasma. The method employed a simple liquid-liquid extraction of etoricoxib, salicylic acid, valdecoxib, ketoprofen, nimesulide and celecoxib and internal standard (IS, DRF-4367) from human plasma (500 microL) into acetonitirile. The organic layer was separated and evaporated under a gentle stream of nitrogen at 40 degrees C. The residue was reconstituted in the mobile phase and injected onto a Kromasil KR 100-5C18 column (4.6 x 250 mm, 5 microm). The chromatographic separation was achieved by gradient elution consisting of 0.05 M formic acid (pH 3)-acetonitrile-methanol-water at a flow rate of 1.0 mL/min. The eluate was monitored using an ultraviolet (UV) detector set at 235 nm. The ratio of peak area of each analyte to IS was used for quantification of plasma samples. Nominal retention times of etoricoxib, salicylic acid, valdecoxib, ketoprofen, nimesulide, IS and celecoxib were 15.63, 17.20, 21.66, 24.95, 26.27, 30.24 and 32.22 min, respectively. The standard curve for etoricoxib, salicylic acid, valdecoxib, ketoprofen and celecoxib was linear (r2 > 0.999) in the concentration range 0.1-50 microg/mL and for nimesulide (r2 > 0.999) in the concentration range 0.5-50 microg/mL. Absolute recovery was >83% from human plasma for all the analytes and IS. The lower limit of quantification (LLOQ) of nimesulide was 0.5 microg/mL and for etoricoxib, salicylic acid, valdecoxib, ketoprofen and celecoxib the LLOQ was 0.1 microg/mL. The inter- and intra-day precisions in the measurement of QC samples, 0.1, 0.3, 15.0 and 40.0 microg/mL (for all analytes except nimesulide), were in the range 2.29-9.37% relative standard deviation (RSD) and 0.69-10.28% RSD, respectively. For nimesulide the inter- and intra-day precisions in the measurement of quality control (QC) samples, 0.5, 1.5, 15.0 and 40.0 microg/mL, were in the range 3.21-7.37% RSD and 0.97-7.06% RSD, respectively. Accuracy in the measurement of QC samples for all analytes was in the range 91.03-106.38% of the nominal values. All analytes including IS were stable in the battery of stability studies, viz. bench top, autosampler and freeze-thaw cycles. Stability of all analytes was established for 21 days at -20 degrees C. The application of the assay in an oral pharmacokinetic study in rats co-administered with celecoxib and valdecoxib is described.

Active articulation for future space applications inspired by the hydraulic system of spiders.

This paper presents and discusses a novel mechanism which was conceived taking inspiration from the micro-hydraulic system used by spiders to extend their legs. The mechanism has the potential to be used in future space applications, although the harsh space conditions, and in particular outgassing, should be carefully addressed in the design of a space-qualified model. The new system has one degree of freedom and is actuated by a pressurized fluidic system. The prototype, which has been designed, simulated, built and tested, is of compact size and presents a repeatable behaviour. The relation between pressure and rotation is approximately linear. The mechanism is suitable for a modular configuration in which several elastic joint modules are joined together. This modular configuration allows large rotations and does not increase the complexity of the actuation. A single module bends about 1.8 degrees when the pressure of the working fluid is 1.2 MPa.

Vascularity and uptake of photosensitizer in small human tumor nodules: implications for intraperitoneal photodynamic therapy.

PURPOSE: i.p. spread of cancers is a common clinical problem, with limited treatment options leading to morbidity and death. i.p. photodynamic therapy (IP-PDT) combines maximal surgical debulking of gross tumor with intraoperative light delivery to the peritoneum after preoperative i.v. injection of photosensitizer to treat residual disease. An issue of concern in IP-PDT is the potential lack of photosensitizer uptake by residual small tumor nodules (STNs) < or =5 mm in maximum diameter and by microscopic residual disease caused by incomplete development of a vascular supply. This study examined the existence of vasculature and Photofrin (PF) uptake in STNs in 12 patients in a Phase II clinical trial for IP-PDT. EXPERIMENTAL DESIGN: Patients received PF 2.5 mg/kg i.v. 48 h before surgery. STNs obtained during surgery were cryosectioned, immunostained for platelet/endothelial cell adhesion molecule 1, and analyzed by light microscopy. Mean vascular densities in STNs were determined by counting microvessels within a x200 field (0.28 mm(2) area). Sections were also examined for PF uptake by fluorescence image analysis using an epifluorescence microscope and IPLab Spectrum software. RESULTS: Data obtained showed that tumors as small as 1 mm in diameter stained positive for platelet/endothelial cell adhesion molecule 1 and contained PF. A negative control from a patient not given PF showed no detectable fluorescence. The average of all mean vascular densities in STNs was determined to be 100 +/- 29. CONCLUSIONS: We conclude that STNs, as small as 1 mm in diameter, have a functional vasculature, because these tumors show PF uptake after i.v. delivery. Both properties are crucial for the treatment of residual STNs by IP-PDT after surgical debulking.

Estrogen modulates the expression of myosin heavy chain in detrusor smooth muscle.

The effect of low serum estrogen levels on urinary bladder function remains poorly understood. Using a rabbit model, we analyzed the effects of estrogen on the expression of the isoforms of myosin, the molecular motor for muscle contraction, in detrusor smooth muscle. Expression of myosin heavy chain (MHC) isoforms, which differ in the COOH-terminal (SM1 and SM2) and the NH(2)-terminal (SM-A and SM-B) regions as a result of alternative splicing of the mRNA at either the 3'- or 5'-ends, was analyzed in age-matched female rabbits that were sham operated, ovariectomized (Ovx), and given estrogen after ovariectomy (4 rabbits/group). Ovx rabbits showed a significant decrease in the overall MHC content per gram of wet detrusor smooth muscle compared with controls (P < 0.04), which was reversed by estrogen replacement (P < 0.02). MHC content, as a proportion of total milligram of protein in the bladder tissue extracted, was also increased in estrogen-treated Ovx rabbits. Quantitative competitive RT-PCR revealed 1.72-, 2.63-, and 5.82 x 10(6) copies of MHC mRNA/100 ng total mRNA in Ovx, control, and estrogen-treated rabbits, respectively (P < 0.01). RT-PCR analysis using oligonucleotides specific for the region containing the SM1/SM2 MHC alternative splice sites indicated a lower SM2-to-SM1 ratio in estrogen-treated compared with control and Ovx rabbits (P < 0.05). Similarly, SDS-PAGE analysis of extracted myosin from estrogen-treated rabbits revealed a significantly lower SM2-to-SM1 isoform ratio compared with control and Ovx rabbits (P < 0.05). Expression of the SM-A and SM-B isoforms was not affected. These results indicate that myosin content is increased upon estrogen replacement in Ovx rabbits and that the abundance of SM1 relative to SM2 is greater in estrogen-treated rabbits compared with normal and Ovx rabbits. These data suggest that estrogen affects alternative splicing at the 3'-end of the MHC pre-mRNA to increase the proportion of SM1 vs. SM2.

Photofrin-mediated photodynamic therapy induces vascular occlusion and apoptosis in a human sarcoma xenograft model.

Photodynamic therapy (PDT) involves light activation of a photosensitizer, resulting in oxygen-dependent, free radical-mediated cell death. Little is known about the efficacy of PDT in treating human sarcomas, despite an ongoing clinical trial treating i.p. sarcomatosis. The present study evaluates PDT treatment of a human sarcoma xenograft in nude mice and explores the mechanism of PDT-mediated antitumor effect. Athymic nude mice, 6-8 weeks of age, were s.c. injected with 5 x 10(6) cells of the A673 human sarcoma cell line. Tumors were allowed to grow to a diameter of about 10 mm. Photofrin (PF), 10 mg/kg, was injected by tail vein, and 24 h later, 630 nm light was delivered to the tumor with fluences of 50, 100, 150, or 300 J/cm2 at a fluence rate of 250 mW/cm2. To assess the efficacy of PDT in the treatment of sarcomas, photosensitizer uptake/retention studies and dose-response studies were performed. Studies carried out to determine the mechanism of tumor response included tumor temperature measurements before, during, and after treatment; tumor vascular perfusion studies with laser Doppler; electron microscopic analysis of tumor sections for vascular occlusion; and analysis of tumor cryosections for endothelial cell damage, apoptosis, and necrosis. At all time points of analysis, photosensitizer levels were greater in tumor than in muscle. Dose-response studies showed that at 100 J/cm2, five of six mice had a complete response to treatment, one of six had a partial response, and no deaths occurred. Temperature measurements indicated that thermal injury did not contribute to tumor response. Vascular perfusion studies demonstrated a significant reduction in blood flow as early as 6 h after PDT. Electron micrographs revealed erythrostasis in tumor microvessels starting as early as 2 h after treatment and complete occlusion of blood vessels by 12 h. Starting as early as 4 h after PDT, apoptosis first appeared in endothelial cells lining the occluded blood vessels and became more widespread at later time points. PDT is an effective treatment for this human sarcoma xenograft in nude mice. The mechanism of tumor destruction in this model appears to be vascular damage with initial apoptosis in tumor endothelial cells and delayed tumor cell apoptosis. This therapy may be valuable in the treatment of patients with sarcomatosis.

Expression of myosin isoforms in smooth muscle cells in the corpus cavernosum penis.

Corpus cavernosum smooth muscle (CCSM) in the penis is unique in that it exhibits a high resting tone and, on stimulation, the muscle cells relax, allowing cavernous spaces to fill with blood, which results in an erection (tumescence). During detumescence, the muscle cells contract and return to the state of high resting tone. This study was undertaken to determine whether CCSM with these unique properties contains myosin isoforms typical of aorta or bladder smooth muscles, muscles that exhibit tonic and phasic characteristics, respectively. RT-PCR revealed that normal CCSM contains an SM2/SM1 mRNA ratio of 1.2:1 (similar to the rabbit aorta). Approximately 31% of the myosin heavy chain transcripts possess a 21-nt insert (predominant in bladder smooth muscle but not expressed in aorta) that encodes the seven-amino acid insert near the NH2-terminal ATP binding region in the head portion of the myosin molecule found in SMB, with the remaining mRNA being noninserted (SMA). Quantitative competitive RT-PCR revealed that the CCSM possesses approximately 4.5-fold less SMB than the bladder smooth muscle. Western blot analysis using an antibody specific for the seven-amino acid insert reveals that both SM1 and SM2 in the CCSM contain the seven-amino acid insert. Furthermore, SMB containing the seven-amino acid insert was localized in the CCSM by immunofluorescence microscopy using this highly specific antibody. The analysis of the expression of LC17 isoforms a and b in the CCSM revealed that it is similar to that of bladder smooth muscle. Thus the CCSM possesses an overall myosin isoform composition intermediate between aorta and bladder smooth muscles, which generally express tonic- and phasiclike characteristics, respectively. Two-dimensional gel electrophoresis showed a relatively low level (approximately 10%) of Ca2+-dependent light-chain (LC20) phosphorylation at the basal tone, which reaches approximately 23% in response to maximal stimulation. The presence of noninserted and inserted myosin isoforms with low and high levels of actin-activated ATPase activities, respectively, in the CCSM may contribute to the ability of the CCSM to remain in a state of high resting tone and to relax rapidly for normal penile function.