Tendon arthroplasty for basal fourth and fifth metatarsal arthritis.
Arthritis of the fourth and fifth tarsometatarsal joints, recalcitrant to nonoperative treatment, presents a difficult clinical situation. As part of the lateral rays, these joints have considerable motion, making fusion a very disabling operation. Between 1990 and 1998, 12 patients, who had failed nonoperative treatment, underwent resection arthroplasty of the base of the fifth or fourth and fifth metatarsals with tendon interposition. Preoperative differential injections had confirmed the source of pain in eight cases. Patients were evaluated using the American Orthopaedic Foot and Ankle Society (AOFAS) midfoot scale; a visual analogue scale to assess perception of pain and disability; a satisfaction index; and, where possible, a comprehensive physical examination. At an average of 25 months follow-up, the average AOFAS score was 64.5. On the visual analogue scale, pain improved an average of 35% and disability improved 10%. Six of the eight patients who participated in this study were satisfied with the operation and would undergo the procedure again for similar symptoms. Patients with a higher postoperative score on the AOFAS midfoot rating scale were statistically more likely to have had a positive differential injection preoperatively. We believe a lateral column tarsometatarsal resection arthroplasty is an effective salvage operation when lateral column midfoot arthritis is confirmed by differential injection and nonoperative measures have provided inadequate relief.


SARM1 knockout does not rescue neuromuscular phenotypes in a Charcot-Marie-Tooth disease Type 1A mouse model.
Charcot-Marie-Tooth disease Type 1A (CMT1A) is caused by duplication of the PMP22 gene and is the most common inherited peripheral neuropathy. Although CMT1A is a dysmyelinating peripheral neuropathy, secondary axon degeneration has been suggested to drive functional deficits in patients. Given that SARM1 knockout is a potent inhibitor of the programmed axon degeneration pathway, we asked whether SARM1 knockout rescues neuromuscular phenotypes in CMT1A model (C3-PMP) mice. CMT1A mice were bred with SARM1 knockout mice to generate CMT1A/SARM1-/- mice. A series of behavioral assays were employed to evaluate motor and sensorimotor function. Electrophysiological and histological studies of the tibial branch of the sciatic nerve were performed. Additionally, gastrocnemius and soleus muscle morphology were evaluated histologically. Although clear behavioral and electrophysiological deficits were observed in CMT1A model mice, genetic deletion of SARM1 conferred no significant improvement. Nerve morphometry revealed predominantly myelin deficits in CMT1A model mice and SARM1 knockout yielded no improvement in all nerve morphometry measures. Similarly, muscle morphometry deficits in CMT1A model mice were not improved by SARM1 knockout. Our findings demonstrate that programmed axon degeneration pathway inhibition does not provide therapeutic benefit in C3-PMP CMT1A model mice. Our results indicate that the clinical phenotypes observed in CMT1A mice are likely caused primarily by prolonged dysmyelination, motivate further investigation into mechanisms of dysmyelination in these mice and necessitate the development of improved CMT1A rodent models that recapitulate the secondary axon degeneration observed in patients.


Mitral and tricuspid repair in an adult achondroplastic patient.
Achondroplasia is a rare genetic disorder with multiple anatomic abnormalities making surgery, and anesthesia more challenging. The reported cases of cardiac interventions in this population are few, and to the best of our knowledge, only 16 cases are documented. Herein, we represent the first case of mitral and tricuspid repair in one of these patients, which performed without the need for specific equipment with a smooth postoperative course.


Complete Clinical Response in Stage IVB Endometrioid Endometrial Carcinoma after First-Line Pembrolizumab Therapy: Report of a Case with Isolated Loss of PMS2 Protein.
Endometrial cancer is the only gynecological cancer that is rising in incidence and associated mortality worldwide. Although most cases are diagnosed as early stage disease, with chances of cure after primary surgical treatment, those with advanced or metastatic disease have a poor prognosis because of the quality of treatment options that are currently available. Mismatch repair (MMR)-deficient cancers are susceptible to programmed cell death protein 1 (PD-1)/programmed cell death ligand 1 inhibitors. The US Food and Drug Administration granted accelerated approval to pembrolizumab for MMR-deficient tumors, the first tumor-agnostic approval for a drug. We present a case of stage IV endometrioid endometrial carcinoma with isolated PMS2 protein loss, in which treatment with first-line pembrolizumab therapy achieved a complete clinical and pathological response of tumor.


Frequency organization of the 40-Hz auditory steady-state response in normal hearing and in tinnitus.
We used the 40-Hz auditory steady-state response (SSR) to compare for the first time tonotopic frequency representations in the region of primary auditory cortex (PAC) between subjects with chronic tinnitus and hearing impairment and normal hearing controls. Frequency representations were measured in normal hearing (n=17) and tinnitus (n=28) subjects using eight carrier frequencies between 384 and 6561 Hz, each amplitude modulated (AM) at 40-Hz on trials of 3 min duration under passive attention. In normal hearing subjects, frequency gradients were observed in the medial-lateral, anterior-posterior, and inferior-superior axes, which were consistent with the orientation of Heschl's gyrus and with functional organization revealed by fMRI investigations. The frequency representation in the right hemisphere was approximately 5 mm anterior and approximately 7 mm lateral to that in the left hemisphere, corroborating with MEG measurements hemispheric asymmetries reported by cytoarchitectonic studies of the PAC and by MRI morphometry. In the left hemisphere, frequency gradients were inflected near 2 kHz in normal hearing subjects. These SSR frequency gradients were attenuated in both hemispheres in tinnitus subjects. Dipole power was also elevated in tinnitus, suggesting that more neurons were entrained synchronously by the AM envelope. These findings are consistent with animal experiments reporting altered tonotopy and changes in the response properties of auditory cortical neurons after hearing loss induced by noise exposure. Degraded frequency representations in tinnitus may reflect a loss of intracortical inhibition in deafferented frequency regions of the PAC after hearing injury.


A self-inducible heterologous protein expression system in Escherichia coli.
Escherichia coli is an important experimental, medical and industrial cell factory for recombinant protein production. The inducible lac promoter is one of the most commonly used promoters for heterologous protein expression in E. coli. Isopropyl-β-D-thiogalactoside (IPTG) is currently the most efficient molecular inducer for regulating this promoter's transcriptional activity. However, limitations have been observed in large-scale and microplate production, including toxicity, cost and culture monitoring. Here, we report the novel SILEX (Self-InducibLe Expression) system, which is a convenient, cost-effective alternative that does not require cell density monitoring or IPTG induction. We demonstrate the broad utility of the presented self-inducible method for a panel of diverse proteins produced in large amounts. The SILEX system is compatible with all classical culture media and growth temperatures and allows protein expression modulation. Importantly, the SILEX system is proven to be efficient for protein expression screening on a microplate scale.


Longitudinal changes in microbiology and clinical periodontal parameters after removal of fixed orthodontic appliances.
The aim of this longitudinal study was to monitor patients' microbiological and clinical parameters from bracket placement up to 3 months post-treatment. Twenty-four patients (10 males and 14 females, aged 14.6 ± 1.0 years) were included in this investigation. Microbiology (sub- and supragingival), periodontal probing depth (PPD), bleeding on probing (BOP), and gingival crevicular fluid (GCF) flow were assessed at baseline (T1), at bracket removal (T2), and 3 months post-treatment (T3). A statistical comparison was made over time and between the banded, bonded, and control sites. Repeated measurements on patients were taken into account by modelling the patients as a random factor. Except for PPD and BOP, values were log-transformed before analysis. Corrections for simultaneous hypothesis testing were performed via simulation. The results demonstrated that sub- and supragingival colony-forming units ratio (CFU ratio aerobe/anaerobe) decreased significantly (relatively more anaerobes) at T2 compared with T1. Between T2 and T3 no significant increase in CFU ratio was seen, resulting in a significantly lower CFU ratio at T3 compared with T1 for subgingival plaque. The difference concerning supragingival plaque between T3 and T1 was not significant. Clinical parameters PPD, POB, and GCF flow showed a significant increase between T1 and T2. Between T2 and T3 these variables decreased significantly but remained significantly higher than at T1 [except for BOP values at the bonded sites (P = 0.0646)]. Placement of fixed orthodontic appliances has an influence both on microbial and clinical periodontal parameters, which were only partly normalized, 3 months following the removal of the appliances.


Tomographic parameters for the detection of keratoconus: suggestions for screening and treatment parameters.
Tomographic corneal evaluation has added significantly more information in the evaluation of the ectatic cornea. Additional information from the posterior cornea and a full corneal thickness map in addition to anterior corneal analysis has increased the ability to identify early and subtle corneal changes. Although these newer examination modalities have increased our sensitivity in refractive screening, problems still persist when attempting to assure treatment trials (such as collagen cross-linking) for early disease are performed to an appropriate patient population. This article will review past and present diagnostic capabilities with a particular emphasis on refractive screening and early treatment studies and stress the limitations of our current diagnostic ability when it comes to diagnosing early ectatic change.


Use of pamidronate disodium to reduce cholecalciferol-induced toxicosis in dogs.
To determine whether pamidronate disodium can reduce cholecalciferol-induced toxicosis in a dose-related manner. 20 clinically normal, 8- to 12-month-old male Beagles. All dogs were given 8 mg of cholecalciferol (CCF)/kg of body weight once orally, then were randomly assigned to 4 groups of 5 dogs each. Dogs were treated with IV administration of 0.9% NaCl solution (SC group), 0.65 mg of pamidronate/kg in 0.9% NaCl solution (LP group), 1.3 mg of pamidronate/kg in 0.9% NaCl solution (MP group), or 2.0 mg of pamidronate/kg in 0.9% NaCl solution (HP group) on days 1 and 4 after administration of CCF. Dogs were observed for 14 days, and serial blood samples were collected for serum biochemical, electrolyte, and 25-hydroxyvitamin D3 analyses. Urine samples were collected for determination of specific gravity. Glomerular filtration rate (GFR) was determined by plasma iohexol clearance. Histologic examination of renal tissue was performed. One dog in the SC group was euthanatized 3 days after administration of CCF because of severe clinical signs of toxicosis. Dogs in the HP group had significantly higher mean GFR (day 3), serum potassium concentrations (day 14), and urine specific gravity (days 7 and 14) and significantly lower mean serum creatinine concentrations and total calcium X phosphorus concentration product (days 4 and 7) than dogs in the SC group. Dogs in the HP group had no abnormal findings on histologic examination of renal tissue, dogs in the LP and MP groups had trace to mild mineralization of renal tissue, and dogs in the SC group had moderate mineralization and cellular necrosis of proximal renal tubules. Pamidronate disodium is a potentially useful drug to reduce CCF-induced toxicosis and other causes of hypercalcemia associated with increased bone resorption in dogs.


Colonization of arbuscular-mycorrhizal fungi on Ri T-DNA transformed roots in synthetic medium.
Hairy root culture of tomato (Lycopersicon esculantum L.) was induced with three strains of Agrobacterium rhizogenes namely A4, ATCC 15834 and LBA 9402. The best response in terms of growth of hairy root was observed with A. rhizogenes strain A4 and LBA 9402 followed by ATCC 15834. Hairy roots were maintained on Murashige and Skoog (MS) medium but it could also grow on minimal (M) medium. Spores of Gigaspora margarita were isolated by wet sieving and decanting method and further recovered by sucrose density gradient method. A new method for surface sterilization of spores has been described which is simpler than the methods described earlier. Surface sterilized spores of G. margarita were used for inoculation of transformed roots grown on M medium as it was found more favourable for germination and growth of spores. During co-cultivation, mycorrhizal spore germination and its penetration into root cortex were observed. Inter and intracellular mycelial spread and formation of arbuscules were also observed in the cortical region of transformed roots of this plant.


Modification and Potential Application of Short-Chain-Length Polyhydroxyalkanoate (SCL-PHA).
As the only kind of naturally-occurring biopolyester synthesized by various microorganisms, polyhydroxyalkanoate (PHA) shows a great market potential in packaging, fiber, biomedical, and other fields due to its biodegradablity, biocompatibility, and renewability. However, the inherent defects of scl-PHA with low 3HV or 4HB content, such as high stereoregularity, slow crystallization rate, and particularly the phenomena of formation of large-size spherulites and secondary crystallization, restrict the processing and stability of scl-PHA, as well as the application of its products. Many efforts have focused on the modification of scl-PHA to improve the mechanical properties and the applicability of obtained scl-PHA products. The modification of structure and property together with the potential applications of scl-PHA are covered in this review to give a comprehensive knowledge on the modification and processing of scl-PHA, including the effects of physical blending, chemical structure design, and processing conditions on the crystallization behaviors, thermal stability, and mechanical properties of scl-PHA.


Meeting the mental health needs of elderly clients.
1. Older individuals in both community and institutional settings have traditionally underused mental health services. Providers of such care devote a minimum of their professional time to those over 65 years of age. 2. Ageism and difficulty with diagnosis are the most frequent explanations for inadequate mental health care to the elderly. This study found that lack of referrals and failure of the aged to seek services partially accounted for underuse. 3. Legislative changes mandating that aged persons be screened for and receive appropriate mental health services will increase the demand for qualified providers. 4. Nursing's approach to care of the older adult, which focuses on both physiological and psychological needs, can facilitate the delivery of comprehensive effective mental health interventions in and out of the institutional setting.


Systematic error in the determination of nocturnal blood pressure dipping status by ambulatory blood pressure monitoring.
Vertical displacement of the arm relative to the heart causes inverse changes in blood pressure of approximately 0.8 mmHg for every centimetre change in arm position. Therefore a potential confounding issue in assessing diurnal variation in blood pressure during ambulatory blood pressure monitoring (ABPM) is arm position during sleep. An increase in the number of patients with 'excessive' nocturnal dipping (> 20% decrease in night/day blood pressure) was observed following the creation of an instructional videotape in which patients were advised to muffle the noise of the monitor with a pillow at night. This raised the possibility that patients were placing their arm on top of the pillow reducing nocturnal blood pressure readings. Ambulatory blood pressure monitoring data from 184 patients prior to and from 193 patients following specific instructions not to put their arm on top of the pillow was examined. Following the instructions, the percentage of patients with 'excessive' nocturnal dipping in blood pressure decreased (excessive systolic dipping 17.4 versus 8.8%, P = 0.014; excessive diastolic dipping 37 versus 24.4%, P = 0.01). Consistent with an increase in the ratio of nocturnal/day pressures, there was an increase in the percentage of patients with inadequate nocturnal dipping (< 10% decrease in night/day blood pressure; systolic dipping 33.7 versus 45.6%, P = 0.02; diastolic dipping 13.0 versus 31.6%, P < 0.001) Instructing patients to avoid resting their arm on a pillow at night has a substantial effect on the classification of nocturnal dipping status. Patients need clear instructions not to place their arm on a pillow at night during blood pressure monitoring.


Breast feeding and neonatal behavioral state.
18 normal newborn infants were studied at the age of 5-6 days. Each infant was given breast or bottlefeeding in 2 different sessions on the same day. The newborn assessment was also done in these 2 3-hour sessions, in which sucking and behavioral state were determined. 10 of 18 infants were found to fall asleep soon after the breastfeeding, and to have more quiet sleep and less waking than after bottlefeeding. All of the mothers showed good breast-milk secretion when examined. The other 8 infants, who were not given enough milk, did not show the abovementioned tendency. These results suggest that breastfeeding exerts good influences on neonatal behavior.


[Expression of ezrin in human lung carcinoma and its clinicopathologic significance.].
Metastasis is the main cause of cancer related mortality, and identification of genes involved in tumor metastasis is important for effective therapies. The membrane cytoskeletal crosslinker participated in several functions including cell adhersion, motility and cell survival, and there is increasing evidence that it regulates tumor progression. However, the role played by ezrin in lung cancer metastasis has not been clearly delineated. The aims of this study are to investigate the ezrin expression pattern in human lung carcinoma and the correlation with clinicopathologic characteristics. Ezrin expression was detected by two- step immunohistochemical staining technique in tumor tissues from 75 lung cancer cases and in normal lung tissues from 16 cases with benign disease and analyzed by the lung cancer clinicopathologic characteristics. The gene and protein level expression of Ezrin in lung cancer cell lines was also detected by Confocal Laser Scanning Microscope (CLSM) and RT-PCR. The positive incidence of ezrin expression (77.3%) was significantly lower in lung cancer tissues than that in normal tissues (100%) (P<0.05), and the down-regulated of ezrin expression was significantly correlated with lymph node metastasis and distant metastasis (P<0.05) and was not correlated with gender, age, tumor size, pathological type, the degree of differentiation of tissue and clinical stage. Otherwise, the subcellular redistribution of ezrin from cell membrane to cell plasma was significantly correlated with lymph node metastasis (P< 0.05), in consistance with in vitro experimental result. High metastasis cell line BE1 had the lowest expression of ezrin under CLSM, similar to RT-PCR, and ezrin was mainly located in cell plasma under CLSM. Ezrin expression is down-regulated in lung cancer tissue and lung cancer cell line, and the locational change from cell membrane to cell plasma may be associated with the oncogenesis and development of lung cancer.


Mechanisms of eye gaze perception during infancy.
Previous work has shown that infants are sensitive to the direction of gaze of another's face, and that gaze direction can cue attention. The present study replicates and extends results on the ERP correlates of gaze processing in 4-month-olds. In two experiments, we recorded ERPs while 4-month-olds viewed direct and averted gaze within the context of averted and inverted heads. Our results support the previous finding that cortical processing of faces in infants is enhanced when accompanied by direct gaze. However, this effect is only found when eyes are presented within the context of an upright face.


A Comprehensive Review on DNA Gyrase Inhibitors.
The newly emerging infectious organisms, the global crisis in antibiotic resistance, and the threat of bioterrorism create an urgent need to discover novel antimicrobial agents. In order to develop novel antimicrobial agents, the mechanism of infectious disease must be better understood. DNA Gyrase is a bacterial enzyme that plays an important role in the replication of DNA and transcription process. It is not present in higher eukaryotes making it a perfect target for developing new antibacterial agents. This review describes the role of DNA gyrase inhibitors in preventing various diseases. In this review, we outline the synthesis and pharmacological action of various novel DNA gyrase inhibitors. DNA gyrase inhibitors were used to treat tuberculosis, bacterial, fungal infections and malaria. DNA gyrase inhibitors mainly act by preventing the supercoiling of DNA strands..


Surgical treatment for an invasive leiomyosarcoma of the inferior vena cava.
A 49-year-old woman presented with right lumbar pain and edema in both legs. Computed tomography showed a large low attenuated mass around and in the S7 segment of the liver involving the right kidney and multiple enlarged mesenteric lymph nodes. There were multiple variably sized discrete nodules in both lungs. Cavography showed subtotal occlusion of the inferior vena cava (IVC). She was successfully treated by wide resection and IVC reconstruction with partial cardiopulmonary bypass and metastasectomy.


Involvement of oxidative stress in D-xylose-induced cataractogenesis in cultured rat lenses.
In order to clarify the involvement of oxidative stress in in vitro sugar-induced cataractogenesis, we examined changes in lipid peroxide, reduced glutathione, vitamin E, and water contents following cataractogenesis in rat lenses cultured with 20 mM D-xylose over a period of 24 h. In D-xylose-treated lenses, an apparent opacity appeared at the equator after 6 h of D-xylose treatment, increases in lipid peroxide and water contents and decreases in reduced glutathione and vitamin E contents occurred with the appearance of opacity. We further examined the effect of the treatment of vitamin E in a liposomal form on cataractogenesis and the changes of lipid peroxide, reduced glutathione, vitamin E, water, and xylitol contents in rat lenses cultured with 20 mM D-xylose for 24 h. This vitamin E treatment significantly prevented not only cataractogenesis, but also an increase in lipid peroxide content and a decrease in vitamin E content in the D-xylose-treated lenses. However, the vitamin E treatment had no effect not only on a decrease in reduced glutathione content, but also on increases in water and xylitol contents, which are known to be induced via aldose reductase, in the D-xylose-treated lenses. These results indicate that not only osmotic stress but also oxidative stress should be involved in cataractogenesis in rat lenses cultured with D-xylose and support the involvement of oxidative stress in in vitro sugar-induced cataractogenesis.


Regulation of the glutamate transporter EAAT4 by PIKfyve.
The excitatory amino-acid transporter EAAT4 (SLC1A6), a Na(+),glutamate cotransporter expressed mainly in Purkinje cells, serves to clear glutamate from the synaptic cleft. EAAT4 activity is stimulated by the serum and glucocorticoid inducible kinase SGK1. SGK1-dependent regulation of the Na(+),glucose transporter SGLT1 (SLC5A1) and the creatine transporter CreaT (SLC6A8) has recently been shown to involve the mammalian phosphatidylinositol-3-phosphate-5-kinase PIKfyve (PIP5K3). The present experiments thus explored whether SGK1-dependent EAAT4-regulation similarly involves PIKfyve. In Xenopus oocytes expressing EAAT4, but not in water injected oocytes, glutamate induced a current which was significantly enhanced by coexpression of PIKfyve and SGK1. The glutamate induced current in Xenopus oocytes coexpressing EAAT4 and both, PIKfyve and SGK1, was significantly larger than the current in Xenopus oocytes expressing EAAT4 together with either kinase alone. Coexpression of the inactive SGK1 mutant (K127N)SGK1 did not significantly alter glutamate induced current in EAAT4-expressing Xenopus oocytes and abolished the stimulation of glutamate induced current by coexpression of PIKfyve. The stimulating effect of PIKfyve was abrogated by replacement of the serine with alanine in the SGK consensus sequence ((S318A)PIKfyve). Furthermore, coexpression of (S318A)PIKfyve significantly blunted the stimulating effect of SGK1 on EAAT4 activity. The observations disclose that PIKfyve indeed participates in the regulation of EAAT4.


Neurotrophin-4 modulates the mechanotransducer Cav3.2 T-type calcium current in mice down-hair neurons.
The T-type Ca2+ channel Cav3.2 is expressed in nociceptive and mechanosensitive sensory neurons. The mechanosensitive D-hair (down-hair) neurons, which innervate hair follicles, are characterized by a large-amplitude Cav3.2 T-current involved in the amplification of slow-moving stimuli. The molecules and signalling pathways that regulate T-current expression in mechanoreceptors are unknown. In the present study, we investigated the effects of NT-4 (neurotrophin-4) on Cav3.2 T-current expression in D-hair neurons in vitro. Interruption of the supply of NT-4 with peripheral nerve axotomy induced a non-transcriptional decrease in the T-current amplitude of fluorogold-labelled axotomized sensory neurons. The T-current amplitude was restored by incubation with NT-4. Deletion of NT-4 through genetic ablation resulted in a similar selective loss of the large-amplitude T-current in NT-4-/- sensory neurons, which was rescued by the addition of NT-4. NT-4 had no effect on the T-current in Cav3.2-/- D-hair neurons. Neither the biophysical properties of the T-current nor the transcript expression of Cav3.2 were modified by NT-4. Pharmacological screening of signalling pathways activated under the high-affinity NT-4 receptor TrkB (tropomyosin receptor kinase B) identified a role for PI3K (phosphoinositide 3-kinase) in the potentiation of T-current. The results of the present study demonstrate the post-transcriptional up-regulation of the Cav3.2 T-current through TrkB activation and identify NT-4 as a target-derived factor that regulates the mechanosensitive function of D-hair neurons through expression of the T-current.


General practitioners and sales representatives: Why are we so ambivalent?
Accepting gifts from pharmaceutical sales representatives (sales reps) or meeting them is correlated with excessive, more expensive and sometimes less rational prescribing. French general practitioners (GPs) tend to hold an unfavorable opinion of the pharmaceutical industry, yet the behavior they adopt with sales reps is generally favorable. Until now, no study has sought to explain the reasons for this discrepancy. This study explores GP experiences to better understand their ambivalent behavior. This qualitative descriptive study was based on semi-structured face-to-face interviews with French GPs in the south-east of France. An interpretative phenomenological approach was chosen to explore individual professional practices and to model the phenomenon through in-depth analysis of semi-structured interviews. A general inductive analysis was carried out. Data were analyzed by researchers from different disciplines (psychology, sociology and general practice). Ten GPs were interviewed for an average of 50 minutes. The analysis revealed three forces that combine to motivate GPs to keep meeting sales reps despite their unfavorable opinion of these visits: practical reasons such as the need for a substitute for continuing education; social and cultural reasons such as courtesy towards representatives; and psychological mechanisms such as cognitive dissonance and a hidden curriculum. The GP-representative relationship is complex and involves psychological mechanisms that the medical profession often fails to recognize. GPs use reps as a convenient tool for continuing education, particularly in the setting of a private practice where GPs feel pressed for time. Cognitive dissonance is a well-supported theory in social psychology that explains how a person maintains a behavior while having an unfavorable opinion of it. Since GP meetings with sales reps start during their internship, they could also be considered as part of a hidden curriculum. The strength of this work is to combine medical, social psychological and sociological perspectives with the original interpretative phenomenological approach. When the veil is lifted on individual ambivalence, the questions raised are more social and political than individual.


Are we any closer to beating the biofilm: novel methods of biofilm control.
A multidisciplinary approach to the treatment and management of biofilms has resulted from the growing appreciation of the role that biofilms play in modern medicine. Conventional antimicrobial agents are generally ineffective against biofilms, and as a result novel laboratory-based and clinical strategies have emerged. The purpose of this review is to analyse the recent literature relating to novel treatment strategies targeting the growing spectrum of clinically relevant biofilms. Microscopy and molecular techniques have provided greater insights into identifying the key bacterial and fungal biofilm pathogens. Knowledge of these microorganisms has provided a foundation for the development of specific molecules, often microbial derived, with antimicrobial and/or biofilm disruptive properties, augmenting conventional antibiotics treatments. The validity of some such rationally designed therapeutics has been explored in clinical trials. Biofilms are inherently difficult to treat, and mechanical disruption is the mainstay of clinical management. With scientific progress in molecular microbiology, there is an abundance of newly discovered molecules and pathways, providing novel therapeutic and prophylactic targets.


"My patients are sicker:" using the Pra risk survey for case finding and examining primary care site utilization patterns in a medicare-risk MCO.
The probability of repeat admissions (Pra) survey is a case-finding tool designed to identify elderly persons at risk for hospitalization. Little is known about the ability of the Pra to assess utilization patterns in a managed care setting. To assess the ability of the Pra survey to identify elders at risk for increased healthcare costs and to determine the utility of this tool in predicting mean per member per year (PMPY) claims among primary care sites in a managed care setting. Mean PMPY paid claims, inpatient admissions, and inpatient days were compared for survey respondents with high, medium, and low Pra scores. Linear regression was used to examine the correlation (R2) between the mean Pra score for respondents and the percentage of high Pra score respondents at each primary care site in the HMO and the sites' mean PMPY paid claims. From a single Medicare-risk health maintenance organization (HMO) with approximately 25,000 members and 234 primary care sites in northeastern and central Pennsylvania, we contacted 24,947 enrollees about the Pra survey. A total of 17,484 (70.1%) patients in 159 of 234 (67.9%) primary care sites responded to the survey. Of these, 17,469 (99%) surveys could be scored. The mean Pra score in this population was 0.27 (standard deviation = 0.108). Pra scores were stratified as follows: 4.5%, 24.1%, and 71.4% of respondents scored high (20.5), moderate (> or = 0.3 and <0.5), and low (<0.3), respectively. Patients with high Pra scores had a mean of $12,611 in PMPY claims, versus $6944 and $3038 for moderate and low scores respectively (P < .0001). The R2 between the mean Pra score for respondents at each primary care site and the sites' mean PMPY Medicare-risk claims was 0.042. If survey respondents assigned to 6 specialty care sites (hematology, oncology, rheumatology, endocrinology, nephrology, and pulmonary medicine) are included, the R2 increased to 0.176. The R2 between the percentage of high-risk individuals with a Pra score of 0.5 or higher at each primary care site and the sites' mean PMPY Medicare-risk claims was 0.0005. In this Medicare-risk HMO population, the Pra survey successfully performed 2 functions: (1) the prospective identification of enrollees at risk for increased healthcare utilization and (2) identification of patient factors that accounted for as much as 17.6% of the variation in utilization between primary sites.


Adenovirus DNA polymerase is a phosphoprotein.
Biological activities of many of the eukaryotic DNA replication proteins are modulated by protein phosphorylation. Investigations of the phosphorylation of adenovirus DNA polymerase (AdPol) have been difficult mainly because of its low level of synthesis in adenovirus-infected HeLa cells. However, when AdPol was overproduced using the recombinant vaccinia virus (RV-AdPol) and the baculovirus expression systems, or by a large scale metabolic labeling of adenovirus 2-infected HeLa cells (native AdPol), in vivo phosphorylation of AdPol could be demonstrated. Phosphoamino acid analysis of [32P]AdPol indicated the presence of phosphoserine independent of the source of AdPol. Comparison of tryptic peptide maps of native AdPol and RV-AdPol revealed that the majority of phosphopeptides were common. Fractionation by high performance liquid chromatography and sequencing of one of the major phosphopeptides revealed serine 67 as a site of phosphorylation. Interestingly, this site is located close to the nuclear localization signal of AdPol and has a consensus substrate recognition sequence for histone H1 (cdc2-related) kinases and mitogen-activated protein kinases. Dephosphorylation of AdPol with calf intestinal alkaline phosphatase resulted in significant decrease in its activity in the in vitro DNA replication initiation assay, suggesting that phosphorylation is important for its biological activity.


A subtype of a Pseudomonas aeruginosa cystic fibrosis epidemic strain exhibits enhanced virulence in a murine model of acute respiratory infection.
The Liverpool epidemic strain (LES) of Pseudomonas aeruginosa is a particularly successful cystic fibrosis (CF) pathogen associated with transmissibility, increased patient morbidity, and, unusually, infection of the non-CF parents of a patient with CF. Using assays for virulence-associated exoproducts, biofilm formation, Caenorhabditis elegans killing, and a murine model of acute respiratory infection, we compared the pathogenic behavior of representatives of 4 subtypes of the LES, including LES431, an isolate associated with the infection of a parent without CF. The quorum-sensing-defective lasR mutant LES400 produced less exoproduct and had less C. elegans killing activity than the other LES subtypes, which were represented by LES431, LESB58, and LESB65. LES431 was deficient in biofilm formation, compared with the other LES sub-types. The LES subtypes displayed a range of virulence in the mouse model, with LES431 being by far the most virulent. The genome-sequenced isolate LESB58, effective at establishing infections in a rat model of chronic infection, was the least virulent subtype in the murine acute infection model. LES isolates display widely variable pathogenic characteristics. LES431, associated with transmission to the non-CF parent of a CF patient, represents a "hypervirulent" subtype more adapted to acute infections than chronic infections.


Clinical Application of Noninvasive Prenatal Testing for Sex Chromosome Aneuploidies in Central China.
The relatively high incidence and the clinical symptoms of sex chromosome aneuploidies (SCAs) make prenatal screening of SCAs an attractive option for pregnant women. However, limited studies have assessed the clinical performance of noninvasive prenatal testing (NIPT) for screening SCAs. This study was performed to evaluate the clinical performance of NIPT for SCAs in singleton pregnancies in central China. Noninvasive prenatal testing was performed using next-generation sequencing. Standard Z-score analysis was used to identify fetal SCAs. NIPT-positive results were confirmed by invasive prenatal diagnosis (IPD). A total of 42,164 pregnant women with singleton pregnancies were recruited in this study. They were divided into the following five groups with different clinical indications: with ultrasound soft index abnormalities (9.23%, 3,892/42,164); with advanced maternal age (22.14%, 9,336/42,164); with high risk for maternal serum screening (MSS) (18.35%, 7,738/42,164); with an intermediate risk for MSS (26.6%, 11,215/42,164); and with low risk (23.68%, 9,983/42,164). In all, 223 women had a high risk for SCAs by NIPT with a positive rate of 0.53%. There was no significant difference associated with the five groups in the positive rate. Of all of the positive results, 89 were 45,X (39.91%), 38 were 47,XXX (17.04%), 31 were 46,XY,del(X) (13.90%), 50 were 47,XXY (22.42%,), and 15 were 47,XYY (6.73%). Finally, 147 participants (65.92%) chose to undergo IPD, and 47 cases were confirmed. The combined positive predictive value (PPV) of NIPT for SCA was 31.97% (47/147). PPV was high for 47,XYY (100%, 11/11), moderate for 47,XXX (42.86%, 9/21) and 47,XXY (45.45%, 15/33), but low for 45,X (16.13%, 10/62) and 46,XY,del(X) (10%, 2/20). The termination rates of Turner syndrome and 47,XXY syndrome were higher than 47,XXX and 47,XYY syndromes. In this relatively large cohort, we evaluated the value of NIPT for SCAs. Our data showed that with informed consent and subsequent professional genetical consulting, NIPT can be a useful method to screen SCAs.


Engagement in risky behaviours by 15-19-year-olds from Polish urban and rural areas.
Anti-health behaviours pose a threat to the health status of the adolescent population in Poland. Among other things, the use and abuse of tobacco, alcohol and psychoactive substances are very dangerous as they may give rise to further health inequalities in young people. The aim of the research was to compare scopes of anti-health behaviours of adolescents in rural and urban areas in Poland. The study consisted of 1,580 adolescents (aged 15-19 years) – 596 from urban areas and 984 from rural areas of the Zachodniopomorskie Province in Poland. 25.61% of 15-19-year-old city dwellers and 30.57% of their rural peers are active tobacco smokers. The smoking habit is more popular among female adolescents than among their male peers. 4.64% of rural 15-19-year-olds, and 4.52% of their city peers, are addicted to narcotics, 11.16% of the surveyed rural adolescents and 6.7% of their peers living in urban areas are addicted to alcohol. More than 50% of adolescents drink alcohol occasionally. A significant number of young people in both rural and urban areas are engaged in risky health behaviours. The frequency of such behaviours depends on the age, gender and place of residence. These disproportions may contribute to future health inequalities in rural and urban areas.


Modern biomarkers in prostate cancer diagnosis.
The most common malignant neoplasm of the urinary tract is prostate cancer (PCa), which is a heterogeneous disease, ranging from very slowly developing and slightly benign to progressing, aggressive, metastatic and fatal, even when properly treated. Existing, imperfect diagnostic methods often lead to over-diagnosis and over-treatment of PCa. That is why new, better PCa biomarkers are being developed. This review summarizes the current results of the most promising and clinically used PCa biomarkers, as well as having the potential to create new diagnostic and prognostic tools, based on the Web of Science (www.apps.webofknowledge.com) and Scopus (www.scopus) databases. com). Limited specificity of the prostate-specific antigen (PSA) test brings a need to develop new and better diagnostic tools. In the last few years, new approaches for providing significantly better biomarkers, an alternative to PSA, have been introduced. Modern biomarkers show improvement in being used as not only a diagnostic procedure, but also for staging, evaluating aggressiveness and managing the therapeutic process. We describe the methods recommended in the diagnosis of PCa and new PCa molecular diagnostics technologies. Individual biomarkers are used in various stages of the PCa diagnostic process, which was presented on the developed diagnostic flowchart describing the role of biomarkers in prostate cancer management. Given the diverse nature of PCa, one diagnostic test will not answer all questions, so the use of several diagnostic methods will allow physicians to provide patients with better, personalized clinical advice.


Error in estimates of tissue material properties from shear wave dispersion ultrasound vibrometry.
Shear wave velocity measurements are used in elasticity imaging to find the shear elasticity and viscosity of tissue. A technique called shear wave dispersion ultrasound vibrometry (SDUV) has been introduced to use the dispersive nature of shear wave velocity to locally estimate the material properties of tissue. Shear waves are created using a multifrequency ultrasound radiation force, and the propagating shear waves are measured a few millimeters away from the excitation point. The shear wave velocity is measured using a repetitive pulse-echo method and Kalman filtering to find the phase of the harmonic shear wave at 2 different locations. A viscoelastic Voigt model and the shear wave velocity measurements at different frequencies are used to find the shear elasticity (mu(1)) and viscosity (mu(2)) of the tissue. The purpose of this paper is to report the accuracy of the SDUV method over a range of different values of mu(1) and mu(2). A motion detection model of a vibrating scattering medium was used to analyze measurement errors of vibration phase in a scattering medium. To assess the accuracy of the SDUV method, we modeled the effects of phase errors on estimates of shear wave velocity and material properties while varying parameters such as shear stiffness and viscosity, shear wave amplitude, the distance between shear wave measurements (delta r), signal-to-noise ratio (SNR) of the ultrasound pulse-echo method, and the frequency range of the measurements. We performed an experiment in a section of porcine muscle to evaluate variation of the aforementioned parameters on the estimated shear wave velocity and material property measurements and to validate the error prediction model. The model showed that errors in the shear wave velocity and material property estimates were minimized by maximizing shear wave amplitude, pulse-echo SNR, delta r, and the bandwidth used for shear wave measurements. The experimental model showed optimum performance could be obtained for delta r = 3 - 6 mm, SNR =35 dB, with a frequency range of 100 to 600 Hz, and with a shear wave amplitude on the order of a few microns down to 0.5 microm. The model provides a basis to explore different parameters related to implementation of the SDUV method. The experiment confirmed conclusions made by the model, and the results can be used for optimization of SDUV.


Pedobarometric evaluation of the normal adult male foot.
Static and dynamic pedobarometric evaluations were performed on the feet of 100 normal adult white men aged from 20 to 49 years old (mean = 29.9 +/- 6.9), using version 3.848 of the F-SCAN system. All evaluations were performed using new pressure sensor insoles with standardized conditions. Maximum vertical forces and plantar peak pressure measurements were taken during 7.88 seconds each of walking in a straight line at subject's own pace and standing. Feet were separated based on their side and lower limb dominance. The means of three consecutive and three alternate tests provided quantitative data. Maximum static and vertical dynamic forces were found to be greater on the dominant side and were significantly correlated with body weight. There were significant differences between dominant and non-dominant sides in static plantar peak pressure evaluations at the forefoot and midfoot, and in the dynamic evaluations at the midfoot. There was a significant correlation between dynamic plantar peak pressures at the midfoot and body weight.


Objective measurement of activation of rigidity: diagnostic, pathogenetic and therapeutic implications in parkinsonism.
1. Quantification of the effect on rigidity of its 'activation', by isometric grip, of standardized pressure, of the contralateral hand, was explored. Torque required to move the forearm through a fixed angle of 40 degrees, at a controlled rate of 0.5 Hz, in a horizontal plane about a pivotal axis aligned to the elbow joint, was recorded before (12 'baseline' recordings), during (10), and after (> or = 8) activation. Work required per unit displacement was calculated. 2. Specificity: Pilot serial daytime measurements gave an overall mean ratio, work required on activation over baseline, of 2.94 (95% CI 2.53, 3.42) in two elderly untreated parkinsonians, and 3.19 (2.75, 3.71) in two elderly subjects with isolated, clinically activation phenomenon, compared with 1.90 (1.64, 2.21) in two elderly without (P < 0.001), whilst two young adults did not activate, 0.98 (0.85, 1.14). In elderly subjects, work required under activation decreased during the day in health (-10 (-5, -14)% h-1, P = 0.0002), showed no significant change in those with clinical activation (4 (-1, 9)% h-1), and increased in parkinsonians (6 (0, 12)% h-1, P = 0.05): there appeared to be a transitionary state. 3. Validation of methodology: Quantifying the same work ratio on a single occasion in 20 aged parkinsonians (P), their spouses (Ps), 20 index controls (C) without parkinsonism, matched to (P), and their spouses (Cs) gave corroborative evidence of a pre-clinical state, defined by other measurements, in the spouses of sufferers. Values for C, Cs and Ps, 1.89 (1.42, 2.52), 2.38 (1.79, 3.17) and 2.93 (2.20, 3.90) respectively, were in consecutive positions, from health to (P, 2.96 (2.22, 3.95)) disease (P = 0.001 for Ps c.f. C; P = 0.1 for Ps c.f. Cs). Data on change over the day may enhance discrimination. 4. Sensitivity to medicines was illustrated, in two parkinsonians, by randomised, placebo balanced and controlled challenges: 1 and 2 tablets, Sinemet CR (Du Pont Pharmaceuticals, each levodopa 200 mg/carbidopa 50 mg) and 1 tablet, Sinemet-Plus (levodopa 100 mg/carbidopa 25 mg), then two 2 mg tablets, benzhexol. The dopaminergic effect (P < 0.001) was selective for activation (treatment.test-condition interaction, P = 0.004), and showed the expected time profiles. The effect of benzhexol (P = 0.008) lacked such selectivity. Its onset (> 4, < or = 6 h) was delayed, compatible with a gastrointestinal anti-muscarinic action and the subjects' ages. 5. Reliability (Fleiss's criterion) was shown to be good in 30 untreated parkinsonians.


The donor lung: infectious and pathologic factors affecting outcome in lung transplantation.
The prevalence of posttransplantation pulmonary infection and the importance of this complication with respect to morbidity in patients undergoing lung transplantation is significant. Over a 1-year period, case histories of all patients undergoing lung transplantation at Barnes Hospital, Washington University, were reviewed to examine the importance of organisms isolated in the donor lung in the development of subsequent invasive infection in transplant recipients. Twenty-eight of 29 bronchial washings (97%) taken from donors before retrieval grew at least one organism. The most common organisms identified were Staphylococcus and Enterobacter. In 12 of these cases (43%), similar organisms were isolated from the tracheobronchial tree of the recipients, and 6 of these recipients (21%) subsequently had invasive pulmonary infections as a result of the organism originally isolated in the donor. We recommend that antibiotic coverage in transplant recipients should be initiated on the basis of Gram stain results and modified on the basis of cultures obtained from the donor lungs. Pathologic analysis of donor lung tissue taken before transplantation was available in 12 cases. Four donors had histologic evidence of established pneumonia in the donor lung, and infections then developed in the recipients. One other patient who received a lung that had widespread bone marrow emboli and subsequent infarction later had a complete anastomotic dehiscence. An additional patient had profound early donor lung dysfunction without any evidence of rejection or infection. Pathologic findings from the donor in this case demonstrated preexistent acute vasculitis with emboli. We suggest that as preservation techniques improve, the opportunities for closer scrutiny of donor lung tissue before implantation will become increasing desirable and feasible.


Technical concepts on blood pool phase SPECT (acquisition, reconstruction).
Guidelines for bone scintigraphy are well established and recommend the use of planar early phase images to investigate a number of clinical indications. With recent advances in gamma camera technology the use of SPECT/CT imaging in the early phases is now possible, offering the potential of improved diagnostic confidence and prognostic value. To date little work has been carried out to optimize the acquisition of early phase bone images using SPECT/CT with most of the available studies acquiring SPECT images after the traditional planar images to allow comparison of the two techniques. Imaging durations of 7 to 10 minutes have been commonly used. However, the use of iterative reconstruction algorithms has been investigated with rapid SPECT imaging to allow imaging durations as low as 4 minutes. The use of CZT based systems with increased sensitivity and improved energy and spatial resolution also offers the potential to reduce imaging times. The optimization of projection measurement order has been investigated as a method of reducing image artefacts as a result of changing tracer distribution during the SPECT acquisition. In this article we consider the current state of early phase SPECT imaging and possible areas for future investigation as well as recommendations for departments looking to adopt blood pool SPECT imaging as part of their routine clinical practice.


Adaptation and preadaptation of Salmonella enterica to Bile.
Bile possesses antibacterial activity because bile salts disrupt membranes, denature proteins, and damage DNA. This study describes mechanisms employed by the bacterium Salmonella enterica to survive bile. Sublethal concentrations of the bile salt sodium deoxycholate (DOC) adapt Salmonella to survive lethal concentrations of bile. Adaptation seems to be associated to multiple changes in gene expression, which include upregulation of the RpoS-dependent general stress response and other stress responses. The crucial role of the general stress response in adaptation to bile is supported by the observation that RpoS(-) mutants are bile-sensitive. While adaptation to bile involves a response by the bacterial population, individual cells can become bile-resistant without adaptation: plating of a non-adapted S. enterica culture on medium containing a lethal concentration of bile yields bile-resistant colonies at frequencies between 10(-6) and 10(-7) per cell and generation. Fluctuation analysis indicates that such colonies derive from bile-resistant cells present in the previous culture. A fraction of such isolates are stable, indicating that bile resistance can be acquired by mutation. Full genome sequencing of bile-resistant mutants shows that alteration of the lipopolysaccharide transport machinery is a frequent cause of mutational bile resistance. However, selection on lethal concentrations of bile also provides bile-resistant isolates that are not mutants. We propose that such isolates derive from rare cells whose physiological state permitted survival upon encountering bile. This view is supported by single cell analysis of gene expression using a microscope fluidic system: batch cultures of Salmonella contain cells that activate stress response genes in the absence of DOC. This phenomenon underscores the existence of phenotypic heterogeneity in clonal populations of bacteria and may illustrate the adaptive value of gene expression fluctuations.


Leu-enkephalin immunoreactivity in the pig pineal gland.
Leu-enkephalin-positive structures in the pig pineal gland were demonstrated immunohistochemically using mouse monoclonal antibody. The pineal glands were obtained from the newborn, 21-day and 7-month old female pig. The immunopositive nerve fibers were observed in the pineal gland as well as in the epithalamic areas. The leu-enkephalin-immunoreactive nerve fibers (single or forming small bundles) were localized mainly in the proximal part of the pineal and they were scarce in other parts. The localization of the fibers points to a central source of this innervation. The study did not show any age-dependent differences in the distribution and density of leu-ekephalin-positive nerve fibers.


[Leprosy trends following implementation of an intervention project in a state capital in the North of Brazil, 2002-2016].
The project Palmas Free of Leprosy was implemented to improve indicators and deal with the disease, since the capital of Tocantins State is the most hyperendemic state capital in Brazil. This study measures the impact of the project's intervention through trend analysis of the priority indicators in Palmas, from 2002 to 2016. The study was based on an analysis of data from the Brazilian Information System for Notifiable Diseases (SINAN) and reports of applied training courses with problem-solving methodology. The indicators for new leprosy cases in Palmas residents were investigated, and the trends were identified by joinpoint regression analysis to assess the results. In the year the intervention project was implemented (2016), the detection rate for new cases in the overall population was 236.3/100,000 inhabitants, and this indicator showed a significant decrease of -7.5% from 2002 to 2014. From 2014 to 2016, there was a significant increase of 104.6% in overall detection. The detection rate in individuals under 15 years of age also showed a reduction of -4.6%, but in the years 2014, 2015, and 2016 there was an increase of 111.1%, together with detection rates for grades 0, 1, and 2, with 59.3%, 225.2%, and 121.7%, respectively. The proportion of cases detected by contact assessment showed a significant increase of 201.1% from 2014 to 2016. The data proved the effectiveness and potentiality of the project's intervention strategy for the diagnosis and control of leprosy in Palmas. The study provided evidence that timely diagnosis by primary care services results in indicators that reflect the real incidence of cases.


DbpA: a DEAD box protein specifically activated by 23s rRNA.
The Escherichia coli protein DbpA is a member of the 'DEAD box' family of putative RNA-dependent ATPases and RNA helicases, so called because they share the highly conserved motif Asp-Glu-Ala-Asp, together with several other conserved elements. We have investigated DbpA expression under conditions where an endogenous promoter is used. In this context, translation initiation does not occur at the previously identified AUG, but at an upstream, in-frame GUG. Mutation of the GUG initiation codon to AUG virtually abolishes DbpA expression, suggesting an unusual translation initiation mechanism. Using an inducible overexpression plasmid, we have purified milligram quantities of DbpA to homogeneity and shown that the purified protein hydrolyses ATP in an RNA-dependent manner. This ATPase activity is interesting in that, unlike that of other DEAD box proteins investigated to date, it absolutely requires a specific bacterial RNA, which we have identified as 23S rRNA. This observation is particularly significant since DbpA will bind other RNAs and DNA, but will only hydrolyse ATP in the presence of 23S rRNA.


Morphology of the temporomandibular joint in skeletal class iii symmetrical and asymmetrical cases: a study by cephalometric laminography.
The aetiology of asymmetric growth in the mandible is not well understood. Previous studies have indicated that the functional lateral shift of the mandible in the period of prepubertal growth may translate to a true skeletal asymmetry, exclusively in skeletal Class III malocclusion. This asymmetry develops more characteristic features during the pubertal and post-pubertal growth periods. Early correction of a functional lateral shift of the mandible is recommended. The purpose of this study was to examine the relationship between the morphology of the temporomandibular joints and asymmetry in skeletal Class III malocclusion in adult female patients. Cephalometric and laminographic findings in 36 asymmetric skeletal Class III patients with a lateral shift of mandible (group 3) were compared to those of 25 symmetric skeletal Class I patients (group 1) and the same number of symmetric skeletal Class III malocclusions (group 2). All the patients had received no orthodontic treatment. The results showed that the TMJ of the side to which the mandible shifted showed a significantly narrower and shorter shape of the condyle head, smaller superior condylar space, and steeper eminence than those of the unshifted side.


Increased radioresistance of EJras-transformed human osteosarcoma cells and its modulation by lovastatin, an inhibitor of p21ras isoprenylation.
Alterations in ras oncogene expression have been associated with increased cellular resistance to ionizing radiation. As an extension of studies with murine cell models, we have now explored the radioresponses of human osteosarcoma (HOS) sub-clones that differ in their EJras expression. Quantitative analysis revealed a tight correlation between the amounts of ras-encoded mRNA and p21 produced, and the degree of cell radioresistance. Interestingly, treatment of the ras-transformed cells with lovastatin, an inhibitor of p21ras post-translational processing via the mevalonate pathway, markedly decreased their radioresistance. Under the experimental conditions used, lovastatin prevented the membrane association, but not the biosynthesis, of p21. The decline in radiation resistance following lovastatin treatment could not be attributed to perturbation of cholesterol metabolism or to non-specific cell-cycle effects. In agreement, lovastatin did not alter the radiation responses of control HOS cells that do not express EJras, or those with an activated met oncogene. The results indicate that elevation in ras gene expression can lead to increased radioresistance of human tumor cells. It appears, however, that p21ras membrane localization is critical for maintenance of the radioresistant phenotype, thus providing a target for pharmacological intervention.


The role of ubiquitination and deubiquitination in the regulation of cell junctions.
Maintenance of cell junctions plays a crucial role in the regulation of cellular functions including cell proliferation, permeability, and cell death. Disruption of cell junctions is implicated in a variety of human disorders, such as inflammatory diseases and cancers. Understanding molecular regulation of cell junctions is important for development of therapeutic strategies for intervention of human diseases. Ubiquitination is an important type of post-translational modification that primarily regulates endogenous protein stability, receptor internalization, enzyme activity, and protein-protein interactions. Ubiquitination is tightly regulated by ubiquitin E3 ligases and can be reversed by deubiquitinating enzymes. Recent studies have been focusing on investigating the effect of protein stability in the regulation of cell-cell junctions. Ubiquitination and degradation of cadherins, claudins, and their interacting proteins are implicated in epithelial and endothelial barrier disruption. Recent studies have revealed that ubiquitination is involved in regulation of Rho GTPases' biological activities. Taken together these studies, ubiquitination plays a critical role in modulating cell junctions and motility. In this review, we will discuss the effects of ubiquitination and deubiquitination on protein stability and expression of key proteins in the cell-cell junctions, including junction proteins, their interacting proteins, and small Rho GTPases. We provide an overview of protein stability in modulation of epithelial and endothelial barrier integrity and introduce potential future search directions to better understand the effects of ubiquitination on human disorders caused by dysfunction of cell junctions.


Light sheet microscopy for tracking single molecules on the apical surface of living cells.
Single particle tracking is a powerful tool to study single molecule dynamics in living biological samples. However, current tracking techniques, which are based mainly on epifluorescence, confocal, or TIRF microscopy, have difficulties in tracking single molecules on the apical surface of a cell. We present here a three-dimensional (3D) single particle tracking technique that is based on prism coupled light-sheet microscopy (PCLSM). This novel design provides a signal-to-noise ratio comparable to confocal microscopy while it has the capability of illuminating at arbitrary depth. We demonstrate tracking of single EGF molcules on the apical surface of live cell membranes from their binding to EGF receptors until they are internalized or photobleached. We found that EGF exhibits multiple diffusion behaviors on live A549 cell membranes. At room temperature, the average diffusion coefficient of EGF on A549 cells was measured to be 0.13 μm(2)/s. Depletion of cellular cholesterol with methyl-β-cyclodextrin leads to a broader distribution of diffusion coefficients and an increase of the average diffusion coefficient at room temperature. This light-sheet based 3D single particle tracking technique solves the technique difficulty of tracking single particles on apical membranes and is able to document the whole "lifetime" of a particle from binding till photobleaching or internalization.


'Short' pancreaticojejunostomy might be a valid option for treatment of chronic pancreatitis in many cases.
The Partington-Rochelle pancreaticojejunostomy (PJ) is an essential management option for patients with chronic pancreatitis (CP) associated with intractable pain and a dilated pancreatic duct (PD). Wide ductotomy and long PJ (L-PJ) have been advocated as the standard of care to ensure full PD decompression. However, the role of short PJ (S-PJ) in a uniformly dilated PD has not yet been evaluated. To evaluate the possible advantages and disadvantages of S-PJ and L-PJ and to interpret the perspective of S-PJ in the treatment of CP. A retrospective review of prospectively collected cohort data was conducted on surgically treated CP patients subjected to side-to-side PJ. The length of the PJ was adapted to anatomical alterations in PD. A comparison was made of S-PJ (< 50 mm) for uniformly dilated PD and L-PJ (50-100 mm) in the setting of multiple PD strictures, calcifications and dilatations. We hypothesized that S-PJ and L-PJ ensure comparable clinical outcomes. The primary outcomes were pain relief and quality of life (QOL); the secondary outcomes were perioperative characteristics, body weight, patients' satisfaction with treatment, and readmission rate due to CP. Overall, 91 patients underwent side-to-side PJ for CP, including S-PJ in 46 patients and L-PJ in 45 patients. S-PJ resulted in better perioperative outcomes: Significantly shorter operative time (107.5 min vs 134 min), lower need for intraoperative (0% vs 15.6%) and total (2.2% vs 31.1%) blood transfusions, and lower rate of perioperative complications (6.5% vs 17.8%). We noted no significant difference in pain relief, improvement in QOL, body weight gain, patients' satisfaction with surgical treatment, or readmission rate due to CP. Based on our data, in the setting of a uniformly dilated PD, S-PJ provides adequate decompression of the PD. As the clinical outcomes following S-PJ are not inferior to those of L-PJ, S-PJ should be preferred as a surgical option in the case of a uniformly dilated PD.


Expression of cyclooxygenase-2 in neoplasms of the mammary gland in bitches.
The aim of the study was to investigate the cyclooxygenase-2 expression in correlation with other neoplasm traits such as: histological type, the differentiation grade, proliferative activity, estrogenic receptor, as well as Hsp70 and p53 proteins expression. Material for the investigation comprised mammary gland tumours, collected from dogs, the patients of veterinary clinics, during surgical procedures. All together 14 adenomas, 66 complex carcinomas, 47 simple carcinomas and 6 solid carcinomas were studied. Evaluations were conducted with histopathological and immunohistochemical methods using suitable antibodies. Expression of COX-2 was observed in 95% of cancers, in case of which, the complex cancers constituted the highest percentage (48.4%). The highest expression of COX-2 was revealed in simple and complex cancers and in cancers with the 3rd degree of histological malignancy. The significant correlation between expression of COX-2 and high mean value of the mitotic index was found. The high expression of COX-2 was also correlated with the expression of protein p53 and expression of the protein Hsp 70. Obtained results suggest that cyclooxygenase-2 may be a prognostic factor, but it requires detailed clinical confirmation.


Infection With a Novel Rickettsiella Species in Emperor Scorpions (Pandinus imperator).
Rickettsiella infection was diagnosed in 4 adult emperor scorpions (Pandinus imperator) from 2 different collections over a 3-year period. One case had a 2-day history of weakness, failure to lift the tail, or respond to stimulation, with rapid progression to death. The other 3 cases were found dead. There were no gross lesions, but histologically the hemolymphatic vasculature and sinuses, presumed hematopoietic organ, heart, midgut and midgut diverticula, nerves, and skeletal muscle were infiltrated with phagocytic and granular hemocytes with necrosis. Phagocytic hemocytes contained abundant intracellular microorganisms that were Fite's acid-fast-positive, Macchiavello-positive, variably gram-positive or gram-negative, and Grocott's methenamine silver-negative. By transmission electron microscopy, hemocytes contained numerous phagocytic vacuoles with small dense bacterial forms (mean 0.603 × 0.163 μm) interspersed with large bacterial forms (mean 1.265 × 0.505 μm) and few intermediary forms with electron-dense nucleoids and membrane-bound crystalline arrays (average 4.72 μm). Transmission electron microscopy findings were consistent with bacteria of the family Coxiellaceae. Based on sequencing the 16S ribosomal RNA gene, the identity was confirmed as Rickettsiella, and phylogenetic analysis of protein-coding genes gidA, rspA, and sucB genes suggested the emperor scorpion pathogen as a new species. This study identifies a novel Rickettsiella causing infection in emperor scorpions and characterizes the unique pathological findings of this disease. We suggest this organism be provisionally named Rickettsiella scorpionisepticum.


Prevalence and therapeutic management of snakebite cases in the health facilities of the Bouenza department from 2009 to 2021, Republic of Congo.
few studies exist of snake bites in the Republic of Congo. This study reports epidemiological and management data on snake bites in the Bouenza department of the Republic of Congo. this is a retrospective and descriptive study based on questionnaire and analysis of files of snakebite victims over a period of 13 years (2009-2021). We collected data on incidence, age, gender, site of the bite, the season of the bite, deaths and treatment. we identified 81 cases of bites recorded in 14 healthcare facilities: 54.32% of cases (44/81) at Nkayi Base Hospital; 11.11% (9/81) at Madingou Base Hospital; 1-5% (1-4/81) at each of the remaining facilities. Eight deaths were recorded in four health facilities. The sex ratio of snakebite victims was 1 (41 males: 40 females). The age most affected was 25-55 years (54.32% or 44/81). The lower limb was the bite site most reported at 13.98% (but in 84% or 68/81 cases the site was not recorded). More bites occurred during the rainy season (80.25%; 65/81). All victims received only symptomatic treatment based on antibiotics, anti-inflammatories and analgesics, as anti-venom serums are unavailable. The recorded incidence of snake bites in Bouenza was 18.62 per 100,000 of population. our study offers a preliminary report from a little-studied region. The incidence of snakebites recorded in Bouenza is lower than expected compared with studies from other African countries, and with earlier (20 years ago) studies from Congo. This may reflect incomplete record-keeping in under-resourced healthcare facilities.


Effect of Salinity on Respiratory Pathways in Root Tips of Tamarix tetragyna.
Oxygen uptake in the presence of exogenous glucose was lower in Tamarix root tips grown in saline media than in those grown in Hoagland solution. This effect was not overcome by raising the external glucose concentration.Glucose uptake and CO(2) evolution were depressed in the presence of NaCl. This effect was observed also when roots were exposed to salinity only during growth but not during uptake. Increasing the external concentration of glucose from 0.01 to 1 mm induced only a 10-fold increase in glucose uptake and CO(2) evolution. However, (14)C evolved in CO(2) as percent of (14)C absorbed, remained constant at all salinity treatments, and was similar at both glucose concentrations.Salinity above 120 mm NaCl increased the percentage of absorbed glucose oxidized via the pentose phosphate pathway, but did not affect the glycolytic pathway. At the same time, salinity depressed the glucose-6-P dehydrogenase, pyruvate kinase, and oxidative phosphorylation. These effects become most evident at a salinity level of about - 10 atm (240 mm), a concentration which is rarely exceeded in the root zone of the natural habitat of the plants.We concluded that Tamarix is reasonably well adapted to the conditions of its habitat, and that salinity affects its root metabolism differently than it does that of pea roots.


Population-Based Prevalence of Oral Conditions as a Basis for Planning Community-Based Interventions: An Epidemiological Study From Rural Burkina Faso.
Objectives: The purpose of the present study was to: (i) provide timely data on dental caries and periodontal conditions in rural Burkina Faso; (ii) identify the oral health practices carried out in this population, and (iii) to prioritize evidence-based interventions at the community level. Methods: Leaning on WHO recommendations for oral health epidemiological studies, clinical examinations and questionnaire-based surveys were conducted in two different age groups (adolescents: 15-19 years old; adults: 35-44 years old) in the health district of Nouna, Burkina Faso. Caries and tooth status were assessed according to the DMFT Index. The periodontal status was assessed by a modified Community Periodontal Index on all teeth present and measuring the level of attachment-loss of six index teeth. Questionnaire items specifically included utilization of oral health care and oral health behaviors. Results: The prevalence of untreated caries was 38% in adolescents and 73% in adults. In terms of periodontal health, 21% of adolescents and 61% of adults had an attachment loss ≥4 mm. Ninety seven percent of adolescents had not attended a dentist in the previous year and 78% of adults had never seen a dentist in their life. About one third of adolescents and adults cleaned their teeth less often than once per day with equal proportions of toothbrushes and traditional chewing sticks made of tree branches. Fluoride toothpaste was used by <10% of study participants. Almost half of the examined people reported to drink a sugar-sweetened beverage at least once a day. Conclusions: Dental caries and periodontitis are highly prevalent in rural Burkina Faso. These findings highlight the key relevance of epidemiological data for identifying people's oral health needs as basis for developing, testing, and implementing oral health interventions and programs. Special emphasis should be put on the design and evaluation of community-based interventions.


[Human immunity system status during 7-day dry immersion].
Subpopulations of lymphocytes, activation potential of T-, B- and NK-cells as well as cytokines production by immunocompetent cells in peripheral blood were studied in five volunteers for 7-day dry immersion without use of countermeasures. Results of the investigation revealed several negative shifts in the immunity and cytokines systems instigated by the experimental conditions. The considerable variability of the immune reactions to immersion suggests individual predisposition to immunological breaks in a changed gravity environment.


Octreotide and octreotate derivatives radiolabeled with yttrium: pharmacokinetics in rats.
Distribution profiles and elimination pathways in rats of two new octreotate derivatives radiolabeled with yttrium, namely Y-DOTAGA-tate and Y-DOTA-t-GA-tate, were compared with those of Y-DOTA-octreotide and Y-DOTA-Tyr(3)-octreotide. All synthetic somatostatin analogues under study were rapidly cleared from the blood and most organs of rats. The main elimination pathway for all peptides under study was urine excretion. High and long-term uptakes of radioactivity in the kidneys and also in organs with high density of somatostatin receptors (the adrenals and pancreas) were found. Radioactivity concentrations in these somatostatin receptor-rich organs were substantially higher for octreotate derivatives in comparison with octreotide analogues; the highest values for Y-DOTAGA-tate were determined. The octreotate derivatives under study appear to be specific ligands for treatment of somatostatin receptor-positive tumors if some mechanism to decrease their kidney retention is provided.


Dexamethasone induces apoptosis in proliferative canine tendon cells and chondrocytes.
Dexamethasone (Dexa) has been commonly used in humans and domestic animals, particularly in the treatment of tendon injuries and cartilage degeneration. However, it is often associated with tendon rupture and impaired tendon and cartilage healing. In the present study, we investigated Dexa's in vitro effects on the growth of cell proliferation and the induction of apoptosis in canine Achilles tendon cells and chondrocytes. Cell proliferation after treatment with Dexa for two to six days was quantified by a 2,3-bis{2-methoxy-4-nitro-5-sulfophenyl}-2H-tetrazolium-5-carboxyanilide inner salt assay (XTT). The results showed that Dexa could inhibit the proliferation of tendon cells and chondrocytes at increasing concentrations (0.1-50 microg/ml) compared with untreated cells. Cell apoptosis was induced by Dexa, as evidenced by the typical nuclear apoptosis using Hoechst 33258 staining. Dexa increased the apoptosis of canine tendon cells and chondrocytes in a time-dependent manner. In canine tendon cells and chondrocytes that were treated with 25 and 50 microg/ml concentration of Dexa, the number of condensed apoptotic nuclei was significantly increased. In addition, culturing with Dexa and the glucocorticoid receptor blocker, mifepristone, significantly arrested apoptosis of tendon cells and chondrocytes. Based on our in vitro data, we hypothesized that in vivo treatment with glucocorticoids may diminish the proliferation of tendon and cartilage cells by increasing apoptosis and suppressing the proliferation. Our findings suggest that Dexa could be used with caution in dogs with articular or tendon problems.


Development of a computer-controlled tensiometer for real-time measurements of tension in tubular organs.
A computer-controlled tensiometer for studying wall tension in tubular organs has been developed. The system consisted of a probe with an inflatable balloon, an impedance planimeter, pressure transducer and amplifier, a pump with RS232 interface and a PC with dedicated software. Circumferential wall tension was computed in real time from pressure and cross-sectional area measurements (tension measurement mode). Wall tension can be maintained on a preset level or be changed as a step or ramp function by a feedback control of the infusion/withdrawal pump (tension control mode). A software regulator adjusted the volume rate (low volume rate when the computed tension was close to the preset value) to minimize overshoot and oscillation. Validation tests were performed and the technique was applied in the human oesophagus. Volume- and tension-controlled balloon distensions elicited secondary peristalsis of increasing intensity that was decreased significantly by the antimuscarinic agent Hyoscine butyl bromide. In tension control mode Hyoscine butyl bromide caused oesophageal relaxation, i.e. CSA to increase and pressure to decay. Furthermore, pronounced pressure relaxation and tension relaxation were observed during volume-controlled distension after administration of Hyoscine butyl bromide.


High Home Blood Pressure Variability Associates With Exaggerated Blood Pressure Response to Cold Stress.
Exaggerated sympathetic cardiovascular (CV) reactivity to stress associates with elevated risk for clinical and preclinical end points of CV disease. It would be useful to identify these individuals, preferably from feasible measurements commonly used in health care. Our study examined the association between home blood pressure (BP) variability and cardiac workload response to whole-body cold exposure. Seventy-five men (55-65 years, 46 hypertensive) measured BP at home twice in the morning and evening for a week. We computed systolic home BP variability as SD of daily means and divided the subjects into groups demonstrating either high or low BP variability. They were exposed to whole-body cold exposure (-10 °C, wind 3 m/second, 15 minutes, winter clothes, standing). BP and heart rate were measured at 3-minute intervals during, and 15 minutes before and after the exposure. Rate-pressure product (RPP) was calculated to represent cardiac workload. Subjects with high systolic home BP variability demonstrated a greater RPP increase in cold conditions compared to those with low BP variability [mean change from baseline (95% CI): 1,850 (1,450 to 2,250) bpm × mm Hg vs. 930 (610, 1,250) bpm × mm Hg, P < 0.01]. This was related to the augmented systolic BP change [31(28, 35) mm Hg vs. 23(20, 26) mm Hg, P < 0.01]. Home BP variability correlated with cold-related RPP (rS = 0.34, P = 0.003) and systolic BP (rS = 0.38, P < 0.001) responses. Moderate whole-body cold exposure increased BP and cardiac workload more among those with higher systolic home BP variability, independently of home BP level. Elevated home BP variability may indicate augmented sympathetically mediated vascular reactivity for environmental stressors. Trial Number NCT02007031.


Nephroprotective Effects of Cardamonin on Renal Ischemia Reperfusion Injury/UUO-Induced Renal Fibrosis.
Acute kidney injury and chronic renal fibrosis are intractable pathological processes to resolve, yet limited strategies are able to effectively address them. Cardamonin (CAD) is a flavonoid with talented antioxidant, anti-inflammatory capacity, and satisfactory biosafety. In our study, animal and cellular models of renal ischemia/reperfusion (I/R) and unilateral ureteral obstruction (UUO) were successfully constructed to confirm whether CAD confers protective effects and underlying mechanisms. Animal experiments demonstrated that CAD application (100 mg/kg) distinctly ameliorated tissue damage and improved renal function. Meanwhile, the continuous oral administration of CAD after UUO surgery efficiently inhibited renal fibrosis as confirmed by hematoxylin-eosin (H&E), Sirius red, and Masson staining as well as the downregulated mRNA and protein expression of collagen I, α-smooth muscle actin (α-SMA), collagen III, and fibronectin. Interestingly, in transforming growth factor β1 (TGF-β1)-stimulated and hypoxia/reoxygenation (H/R)-exposed human kidney-2 (HK-2) cells, protective effects of CAD were again authenticated. Meanwhile, we performed bioinformatics analysis and constructed the "ingredient-target-pathway-disease" network to conclude that the potential mechanisms of CAD protection may be through the regulation of oxidative stress, inflammation, apoptosis, and mitogen-activated protein kinase (MAPK) pathway. Furthermore, experimental data validated that CAD evidently decreased the reactive oxygen species (ROS) production and malondialdehyde (MDA) content while depressing the mRNA and protein expression of inflammatory markers (tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and Il-1β) and inhibiting apoptosis as evidenced by decreased levels of P53, BAX, cleaved caspase-3, and apoptotic rate in renal I/R and UUO models. In addition, the impact of CAD on restraining oxidative stress and inflammation was attributed to its ability to elevate antioxidant enzyme activities including catalase, superoxide dismutase 1 (SOD1), and superoxide dismutase 2 (SOD2) and to inhibit the inflammation-associated MARK/nuclear factor-κB (MAPK/NF-κB) signaling pathway. In conclusion, cardamonin restored the antioxidative capacity to block oxidative stress and suppressed the MAPK/NF-κB signaling pathway to alleviate inflammatory response, thus mitigating I/R-generated acute kidney injury/UUO-induced renal fibrosis in vivo and in vitro, which indicated the potential therapeutic advantage of cardamonin in attenuating acute and chronic kidney injuries.


[Identification of metabolites of imperatorin in rats: based on UHPLC-Q-Exactive Orbitrap MS].
This study aims to identify and analyze the metabolites of imperatorin in rats by UHPLC-Q-Exactive Orbitrap MS. Specifically, after rats were treated(ig) with imperatorin, the plasma, urine, and feces were collected, and the samples were processed by solid phase extraction. Then, UHPLC-Q-Exactive Orbitrap MS was performed. In MS, 0.1% formic acid water(A)-acetonitrile(B) was applied as mobile phase for gradient elution and the data of MS in both positive and negative ion modes were collected. The metabolites of imperatorin in blood, urine, and feces of rats were analyzed to explore the metabolic pathways of imperatorin in rats. According to accurate molecular weight, multistage MS data, MS fragmentation rule of the standard substance, and previous reports, a total of 51 metabolites were identified, with 35, 40, and 16 from plasma, urine, and feces, separately. The main metabolic pathways were oxidization, glucuronidation, isopentenyl removal, sulphation, carboxylation, among others. The conclusion in this study is expected to serve as a reference for the further development and the further pharmacodynamics study of imperatorin.


Activating BRAF mutations in eruptive melanocytic naevi.
Eruptive melanocytic naevi (EMN) are melanocytic proliferations developing rapidly on previously unaffected skin in association with various clinical scenarios, most commonly systemic immunosuppression. However, the exact mechanism leading to development of EMN is not understood. In particular, it is not known whether EMN harbour the BRAF mutations which occur frequently in melanoma and most common naevi. To evaluate whether activating BRAF mutations may play a role in genesis of EMN. Genomic DNA was isolated from 20 EMN from a patient treated with 6-mercaptopurine (6-MP). Primary BRAF genotyping was performed by allelespecific polymerase chain reaction, followed by validation using direct sequencing. The BRAF V600E mutation was identified in 85% of EMN examined. Our results implicate mutational activation of the BRAF–MAPK pathway as a factor in development of EMN in the setting of 6-MP treatment. The mechanism leading to development of EMN in this, and potentially other patients, may relate to synergistic mutagenic effects of thioguanines and ultraviolet (UV) A. Together with the documented importance of BRAF mutations in melanoma development and maintenance, these findings highlight the importance of UVA protection, especially in patients treated with thiopurines such as 6-MP.


Histological diversity in cholangiocellular carcinoma reflects the different cholangiocyte phenotypes.
Cholangiocellular carcinoma (CC) originates from topographically heterogeneous cholangiocytes. The cylindrical mucin-producing cholangiocytes are located in large bile ducts and the cuboidal non-mucin-producing cholangiocytes are located in ductules containing bipotential hepatic progenitor cells (HPCs). We investigated the clinicopathological and molecular features of 85 resected CCs (14 hilar CCs [so-called Klatskin tumor], 71 intrahepatic CCs [ICCs] including 20 cholangiolocellular carcinomas [CLCs], which are thought to originate from HPCs]) and compared these with the different cholangiocyte phenotypes, including HPCs. Immunohistochemistry was performed with biliary/HPC and hepatocytic markers. Gene expression profiling was performed in different tumors and compared with nonneoplastic different cholangiocyte phenotypes obtained by laser microdissection. Invasion and cell proliferation assay were assessed using different types of CC cell lines: KMC-1, KMCH-1, and KMCH-2. Among 51 ICCs, 31 (60.8%) contained only mucin-producing CC features (muc-ICCs), whereas 39.2% displayed histological diversity: focal hepatocytic differentiation and ductular areas (mixed-ICCs). Clinicopathologically, muc-ICCs and hilar CCs showed a predominantly (peri-)hilar location, smaller tumor size, and more lymphatic and perineural invasion compared with mixed-ICCs and CLCs (predominantly peripheral location, larger tumor size, and less lymphatic and perineural invasion). Immunoreactivity was similar in muc-ICCs and hilar CCs and in mixed-ICCs and CLCs. S100P and MUC1 were significantly up-regulated in hilar CCs and muc-ICCs compared with mixed-ICCs and CLCs, whereas NCAM1 and ALB tended to be up-regulated in mixed-ICCs and CLCs compared with other tumors. KMC-1 showed significantly higher invasiveness than KMCH-1 and KMCH-2. Muc-ICCs had a clinicopathological, immunohistochemical, and molecular profile similar to that of hilar CCs (from mucin-producing cholangiocytes), whereas mixed-ICCs had a profile similar to that of CLCs (thought to be of HPC origin), possibly reflecting their respective cells of origin.


Diagnosis of genetic defects through parallel assessment of PLCζ and CAPZA3 in infertile men with history of failed oocyte activation.
Phospholipase C ζ (PLCζ) is considered as a nominee for sperm associated oocyte activating factors and is located back-to-back with CAPZA3, an actin-capping protein controlling actin polymerization during spermiogenesis. They contain a common bidirectional promoter. The objective of this study was to identify individuals with parallel low expression of PLCζ and CAPZA3 mRNA, in hope of detecting genetic defects in this bidirectional promoter. Semen samples were collected from 24 fertile and 59 infertile individuals with total failed, low and high fertilization rate post intra-cytoplasmic sperm injection (ICSI), as well as globozoospermic individuals. Expression of PLCζ and CAPZA3 were assessed by Real time PCR. In addition, PLCζ was assessed by Western blot. Significant correlations between PLCζ with CAPZA3 and also between these two genes with fertilization were observed. Individuals with low fertilization presented significantly lower expression of these two genes. Low expression of PLCζ was also verified by Western analysis. Sequence analysis of bidirectional promoter of these two genes in an individual with parallel low expression of both PLCζ and CAPZA3, revealed a mutation within the CAPZA3 predicted promoter, known as human regulatory factor X4 which is a testis-specific dimeric DNA-binding protein. In the opposite stand, in the same location, the mutation appears to be outside but in the vicinity of PLCζ, in a binding region predicate by Genomatix. Parallel assessment of CAPZA3 with PLCζ at mRNA level in individuals with inability to induce oocyte activation may help researcher to identify genetic defects associated with failed fertilization.


A whole-genome association study for pig reproductive traits.
A whole-genome association study was performed for reproductive traits in commercial sows using the PorcineSNP60 BeadChip and Bayesian statistical methods. The traits included total number born (TNB), number born alive (NBA), number of stillborn (SB), number of mummified foetuses at birth (MUM) and gestation length (GL) in each of the first three parities. We report the associations of informative QTL and the genes within the QTL for each reproductive trait in different parities. These results provide evidence of gene effects having temporal impacts on reproductive traits in different parities. Many QTL identified in this study are new for pig reproductive traits. Around 48% of total genes located in the identified QTL regions were predicted to be involved in placental functions. The genomic regions containing genes important for foetal developmental (e.g. MEF2C) and uterine functions (e.g. PLSCR4) were associated with TNB and NBA in the first two parities. Similarly, QTL in other foetal developmental (e.g. HNRNPD and AHR) and placental (e.g. RELL1 and CD96) genes were associated with SB and MUM in different parities. The QTL with genes related to utero-placental blood flow (e.g. VEGFA) and hematopoiesis (e.g. MAFB) were associated with GL differences among sows in this population. Pathway analyses using genes within QTL identified some modest underlying biological pathways, which are interesting candidates (e.g. the nucleotide metabolism pathway for SB) for pig reproductive traits in different parities. Further validation studies on large populations are warranted to improve our understanding of the complex genetic architecture for pig reproductive traits.


Anatomy of lateral patellar instability: trochlear dysplasia and tibial tubercle-trochlear groove distance is more pronounced in women who dislocate the patella.
A trend toward young women being at greatest risk for primary and recurrent dislocation of the patella is evident in the current literature. However, a causative factor is missing, and differences in the anatomical risk factors between men and women are less defined. To identify differences between the sexes in the anatomy of lateral patellar instability. Case control study; Level of evidence, 3. Knee magnetic resonance images were collected from 100 patients treated for lateral patellar instability. Images were obtained from 157 patients without patellar instability who served as controls. Using 2-way analyses of variance, the influence of patellar dislocation, gender, and their interaction were analyzed with regard to sulcus angle, trochlear depth, trochlear asymmetry, patellar height, and the tibial tubercle-trochlear groove (TT-TG) distance. Mechanisms of injury of first-time dislocations were divided into high-risk, low-risk, and no-risk pivoting activities and direct hits. For all response variables, a significant effect was observed for the incidence of patellar dislocation (all P < .01). In addition, sulcus angle, trochlear asymmetry, and trochlear depth depended significantly on gender (all P < .01) but patellar height did not (P = .13). A significant interaction between patellar dislocation and gender was observed for the TT-TG distance (P = .02). The mean difference in TT-TG distance between study and control groups was 4.1 mm for women (P < .01) and 1.6 mm for men (P = .05). Low-risk and no-risk pivoting injuries were most common in women, whereas first-time dislocations in men occurred mostly during high-risk pivoting activities (P < .01). The data from this study indicate that trochlear dysplasia and the TT-TG distance is more prominent in women who dislocate the patella. Both factors might contribute to an increased risk of lateral patellar instability in the female patient as illustrated by the fact that dislocations occurred most often during low-risk or no-risk pivoting activities in women.


Degradation of nitrobenzene in aqueous solution by ozone-ceramic honeycomb.
The degradation of nitrobenzene by ceramic-honeycomb catalyzed ozonation was investigated. The results showed that the presence of ceramic honeycombs significantly increased the oxidation rate of nitrobenzene by ozone compared to the case of ozone oxidation alone. In this paper, the effects of various factors on the catalytic oxidation were investigated, such as the amount of catalysts, the ozone dosage, the temperature, the pH value and the presence of tert-butanol. With the addition of tert-butanol the removal of nitrobenzene decreased sharply, which appeared to support that, the degradation of nitrobenzene by ozonation followed a radical type mechanism. The EPR experiments verified that higher nitrobenzene removal rate was attributed to more OH* radicals generated in the catalyzed ozonation than ozonation alone.


Spatiotemporal profiles of dental pulp nociception in rat cerebral cortex: an optical imaging study.
Somatosensation is topographically organized in the primary (S1) and secondary somatosensory cortex (S2), which contributes to identify the region receiving sensory inputs. However, it is still unknown how somatosensory inputs from the oral region, especially nociceptive inputs from the teeth, are processed in the somatosensory cortex. We performed in vivo optical imaging and identified the precise cortical regions responding to electrical stimulation of the maxillary and mandibular dental pulp in rats. Electrical stimulation of the mandibular incisor pulp evoked neural excitation in two areas: the most rostroventral part of S1, and the ventral part of S2 caudal to the middle cerebral artery. Maxillary incisor pulp stimulation initially evoked responses only in the ventral part of S2, although later maximum responses were also observed in S1 similar to mandibular incisor stimulation responses. The maxillary and mandibular molar pulp-responding regions were located in the most ventral S2, a part of which was histologically classified as the insular oral region (IOR). In terms of the initial responses, maxillary incisor and molar stimulation induced excitation in the S2/IOR rostral to the mandibular dental pulp-responding region. Contrary to the spatially segregated initial responses, the maximum excitatory areas responding to both incisors and molars in the mandible and maxilla overlapped in S1 and the S2/IOR. Multielectrode extracellular recording supported the characteristic localization of S2/IOR neurons responding to mandibular and maxillary molar pulp stimulation. The discrete and overlapped spatial profiles of initial and maximum responses, respectively, may characterize nociceptive information processing of dental pain in the cortex.


Efficacy of two cleaning and sanitizing combinations on Listeria monocytogenes biofilms formed at low temperature on a variety of materials in the presence of ready-to-eat meat residue.
Biofilms in the food-processing industry are a serious concern due to the potential for contamination of food products, which may lead to decreased food quality and safety. The effect of two detergent and sanitizer combinations on the inactivation of Listeria monocytogenes biofilms was studied. Combination A uses a chlorinated-alkaline, low-phosphate detergent, and dual peracid sanitizer. Combination B uses a solvated-alkaline environmental sanitation product and hypochlorite sanitizer. The survival of bacterial biofilms placed at 4 and 10 degrees C and held for up to 5 days was also addressed. To simulate conditions found in a ready-to-eat meat-processing environment, biofilms were developed in low-nutrient conditions at 10 degrees C (with and without meat and fat residue) on a variety of materials found in a plant setting. Included were two types of stainless steel, three materials for conveyor use, two rubber products, a wall, and floor material. Biofilms developed on all surfaces tested; numbers at day 2 ranged from 3.2 log on silicone rubber to 4.47 log CFU/cm2 on Delrin, an acetal copolymer. Biofilm survival during storage was higher at 4 degrees C (36.3 to 1,621%) than 10 degrees C (4.5 to 83.2%). Small amounts of meat extract, frankfurters, or pork fat reduced biofilm formation initially; with time, the biofilm cell number and survival percentage increased. Cleaning efficacy was surface dependent and decreased with residue-soiled surfaces; biofilms developed on the brick and conveyor material were most resistant. Both detergents significantly (P < 0.05) removed or inactivated biofilm bacteria. The sanitizers further reduced biofilm numbers; however, the reduction was not significant in most cases for the dual peracid. Using a benchmark efficacy of >3-log reduction, combination A was only effective on 50.0% of the samples, Combination B, at 86.1%, was more effective.