Human L- and M-opsins restore M-cone function in a mouse model for human blue cone monochromacy.
Blue cone monochromacy (BCM) is an X-linked congenital vision disorder characterized by complete loss or severely reduced L- and M-cone function. Patients with BCM display poor visual acuity, severely impaired color discrimination, myopia, nystagmus, and minimally detectable cone-mediated electroretinogram. Recent studies of patients with BCM with adaptive optics scanning laser ophthalmoscopy (AOSLO) showed that they have a disrupted cone mosaic with reduced numbers of cones in the fovea that is normally dominated by L- and M-cones. The remaining cones in the fovea have significantly shortened outer segments but retain sufficient structural integrity to serve as potential gene therapy targets. In this study, we tested whether exogenously expressed human L- and M-opsins can rescue M-cone function in an M-opsin knockout (Opn1mw-/- ) mouse model for BCM. Adeno-associated virus type 5 (AAV5) vectors expressing OPN1LW, OPN1MW, or C-terminal tagged OPN1LW-Myc, or OPN1MW-HA driven by a cone-specific promoter were injected subretinally into one eye of Opn1mw-/- mice, while the contralateral eye served as the uninjected control. Expression of cone pigments was determined with western blotting and their cellular localization identified with immunohistochemistry. M-cone function was analyzed with electroretinogram (ERG). Antibodies against cone phototransduction proteins were used to study cone outer segment (OS) morphology in untreated and treated Opn1mw-/- eyes. We showed that cones in the dorsal retina of the Opn1mw-/- mouse do not form outer segments, resembling cones that lack outer segments in the human BCM fovea. We further showed that AAV5-mediated expression of either human M- or L-opsin individually or combined promotes regrowth of cone outer segments and rescues M-cone function in the treated Opn1mw-/- dorsal retina. Exogenously expressed human opsins can regenerate cone outer segments and rescue M-cone function in Opn1mw-/- mice, thus providing a proof-of-concept gene therapy in an animal model of BCM.


Developing minimally invasive procedure quality metrics: one step at a time.
Despite extensive first-hand surgical experience, rank and file members of surgical societies are generally not trained in and have not therefore been included in surgical quality measure development. The purpose of this exercise was to determine if a structured quality metric design tool could bridge this gap, facilitating rapid development of focused quality metrics by minimally invasive surgeon attendees of the April 2018 SAGES Annual Meeting. Expert minimally invasive surgeons attended a 90-min workshop with didactic and interactive quality metric design sessions during the Annual Meeting. The interactive portion was formed around a novel structured quality measure development tool that graded presenting symptoms, short-term complications, and long-term disutility of care. For first-time symptomatic inguinal hernia repair, first-time small to moderate size ventral hernia repair, and elective laparoscopic cholecystectomy, each workgroup was able to develop one quality, one short-term complication, and one long-term disutility metric. A structured quality metric design tool facilitates application of knowledge through rapid development of multifaceted, patient-centric outcomes measures by expert minimally invasive surgeons, otherwise not formally trained in metric development. The exercise also highlighted the need to rigorously define denominator populations and to guard against metric-driven undertreatment.


Outcome of Renal Transplant in Recipients With Vasculitis.
End-stage renal disease develops in a high percentage of patients with vasculitis, in whom kidney transplant has become a therapeutic option. However, limited data are available on the prognosis and outcomes after kidney transplant in these patients. We aimed to compare the long-term graft survival and graft function in 8 renal transplant recipients with vasculitis (granulomatosis with polyangiitis, microscopic polyangiitis, Goodpasture syndrome, and Henoch-Schonlein purpura) with the other kidney recipients at a single center. We conducted a retrospective study of patients followed for chronic renal failure associated with vasculitis before renal transplant. We excluded patients with no biopsy-proven nephropathy. There was no difference in the occurrence of metabolic and cardiovascular complications in our case group compared with the other graft recipients. Infections were frequent and included cytomegalovirus and urinary tract infection. The rates of bacterial and viral infection were equivalent in our population. The incidence of allograft loss was estimated at 1.8%, less than that seen in our entire transplant population. The presence of vasculitis was not significantly related to renal failure (P = .07). Extrarenal relapse occurred in 1 patient with microscopic polyangiitis. Antineutrophil cytoplasmic antibody levels in patients with granulomatosis with polyangiitis and microscopic polyangiitis did not seem to influence the renal outcome (P = .08). Circulating antineutrophil cytoplasmic antibodies were associated with the development of vascular lesions in the graft but were not significantly correlated with graft survival (P = .07). This study supports the theory that renal transplant is an effective treatment option for patients with end-stage renal disease secondary to vasculitis. These patients fare similarly to, if not better than, other patients.


Shaping emotion regulation: attunement, symptomatology, and stress recovery within mother-infant dyads.
The foundations of emotion regulation are organized, in part, through repeated interactions with one's caregiver in infancy. Less is known about how stress physiology covaries between a mother and her infant within these interactions, leaving a gap in our understanding of how the biological basis of emotion regulation develops. This study investigated physiological attunement between mothers and their 5-month-old infants, as well as the influence of maternal depression and anxiety, during stress recovery. During the reengagement phase of the Still Face Paradigm, mother-infant dyads exhibited negative attunement, as measured by inverse covariation of respiratory sinus arrhythmia (RSA). Increases in maternal RSA corresponded to decreases in infant RSA, underscoring dyadic adjustment during recovery. Moreover, infant regulation differed as a function of maternal anxiety, with more anxious mothers having infants with higher RSA during reengagement. Implications for the consolidation of regulatory capabilities within the context of the early caregiving relationship are discussed.


The anion-exchange substrate shuttle process: A new approach to two-stage biomethanation of organic and toxic wastes.
A novel biomethanation process configuration is described which uses improved biocatalysts to enhance the productivity and stability of waste biomethanation systems. The design facilitates the maintenance of acetogenic and methanogenic bacteria under optimum substrate concentrations far above their K(s) values in a two-stage biomethanation system with closed loop reactors. Volatile fatty acid anions (VFA anions) are provided as substrates for the methanogenic stage by using an anion exchange unit coupled to the acidogenic stage. The slow growing and sensitive acetogenic and methanogenic bacteria are protected from oxygen, cationic pollutants, toxins, and microbial contamination by use of the substrate shuttle process. Due to the closed loop process configuration, washout of the ecoengineered biocatalysts is excluded. The modular system configuration allows industrial mass production of the system components. The new biomethanation process enhances both the BTU content of the gas and the methane production over state-of-the-art anaerobic digestor bioreactor concepts.


Influence of Piper betle on hepatic marker enzymes and tissue antioxidant status in D-galactosamine-induced hepatotoxic rats.
D-galactosamine is a well-established hepatotoxicant that induces a diffuse type of liver injury closely resembling human viral hepatitis. D-galactosamine by its property of generating free radicals causes severe damage to the membrane and affects almost all organs of the human body. The leaves of Piper betle L., a commonly used masticatory in Asian countries, possess several biological properties. Our aim is to investigate the in vivo antioxidant potential of P. betle leaf-extract against oxidative stress induced by D-galactosamine intoxication in male albino Wistar rats. Toxicity was induced by an intraperitoneal injection of D-galactosamine, 400 mg/kg body weight (BW) for 21 days. Rats were treated with P. betle extract (200 mg/kg BW) via intragastric intubations. We assessed the activities of liver marker enzymes (aspartate amino-transferase, alanine aminotransferase, alkaline phosphatase, gamma glutamyl transpeptidase) and levels of thiobarbituric acid reactive substances (TBARS), lipid hydroperoxides, superoxide dismutase, catalase, glutathione peroxidase, vitamin C, vitamin E, and reduced glutathione. The extract significantly improved the status of antioxidants and decreased TBARS, hydroperoxides, and liver marker enzymes when compared with the D-galactosamine treated group, demonstrating its hepatoprotective and antioxidant properties.


Effect of particle-hole asymmetry on the Mott-Hubbard metal-insulator transition.
The Mott-Hubbard metal-insulator transition is one of the most important problems in correlated-electron systems. In the past decade, much progress has been made in examining a particle-hole symmetric form of the transition in the Hubbard model with dynamical mean field theory, where it was found that the electronic self-energy develops a pole at the transition. We examine the particle-hole asymmetric metal-insulator transition in the Falicov-Kimball model and find that a number of features change when the noninteracting density of states has a finite bandwidth.


Stick-slip instabilities and shear strain localization in amorphous materials.
We study the impact of strain localization on the stability of frictional slipping in dense amorphous materials. We model the material using shear transformation zone (STZ) theory, a continuum approximation for plastic deformation in amorphous solids. In the STZ model, the internal state is quantified by an effective disorder temperature, and the effective temperature dynamics capture the spontaneous localization of strain. We study the effect of strain localization on stick-slip instabilities by coupling the STZ model to a noninertial spring slider system. We perform a linear stability analysis to generate a phase diagram that connects the small scale physics of strain localization to the macroscopic stability of sliding. Our calculations determine the values of spring stiffness and driving velocity where steady sliding becomes unstable and we confirm our results through numerical integration. We investigate both homogeneous deformation, where no shear band forms, and localized deformation, where a narrow shear band spontaneously forms and accommodates all of the deformation. Our results show that at a given velocity, strain localization leads to unstable frictional sliding at a much larger spring stiffness compared to homogeneous deformation, and that localized deformation cannot be approximated by a homogeneous model with a narrower material. We also find that strain localization provides a physical mechanism for irregular stick-slip cycles in certain parameter ranges. Our results quantitatively connect the internal physics of deformation in amorphous materials to the larger scale frictional dynamics of stick-slip.


QUANTITATIVE VARIATION OF PROGENY FROM CHASMOGAMOUS AND CLEISTOGAMOUS FLOWERS IN THE GRASS DANTHONIA SPICATA.
Progeny from chasmogamous (CH) and cleistogamous (CL) flowers of the grass Danthonia spicata were raised in their native habitat and in the greenhouse in order to determine how genetic variation was distributed among families and between CH and CL progeny within families. Twelve quantitative characters were measured on clones from individuals known to have arisen from either CH or CL flowers on a particular plant. Significant genetic variation existed for all characters measured. Most genetic variation was between families and two morphologically similar groups of families were identified. Relatively little genetic variation was found within families (approximately 5% of the total phenotypic variance). In field-raised plants, variance component analysis suggested that CL progeny were genetically more similar to each other than were CH progeny from the same plant. Levene's test of the average deviation of CH and CL progenies from their group means was nonsignificant but suggested there was a trend (0.05 < P < 0.10) for CH progeny to be more variable than CL progeny in the field but less variable in the greenhouse. The amount and distribution of genetic variation in the study population indicates that selective differentials would be larger among families than within families.


Early development of the gut microbiome and immune-mediated childhood disorders.
The human gastrointestinal tract inhabits a complex microbial ecosystem that plays a vital role in host health through its contributions to nutrient synthesis and digestion, protection from pathogens, and promoting maturation of host innate and adapt immune systems. The development of gut microbiota primarily occurs during infancy and is influenced by multiple factors, including prenatal exposure; gestational age; mode of delivery; feeding type; pre-, pro-, and antibiotic use; and host genetics. In genetically susceptible individuals, changes in the gut microbiota induced by environmental factors may contribute to the development of immune-related disorders in childhood, including atopic diseases, inflammatory bowel disease, irritable bowel syndrome, and necrotizing enterocolitis. Pre- and probiotics may be useful in the prevention and treatment of some immune-related diseases by modulating gut microbiota and regulating host mucosal immune function. The review will discuss recent findings on the environmental factors that influence development of gut microbiota during infancy and its potential impact on some immune-related diseases in childhood. The use of pre- and probiotics for prevention and intervention of several important diseases in early life will also be reviewed.


A comparison of coarse-grained and continuum models for membrane bending in lipid bilayer fusion pores.
To establish the validity of continuum mechanics models quantitatively for the analysis of membrane remodeling processes, we compare the shape and energies of the membrane fusion pore predicted by coarse-grained (MARTINI) and continuum mechanics models. The results at these distinct levels of resolution give surprisingly consistent descriptions for the shape of the fusion pore, and the deviation between the continuum and coarse-grained models becomes notable only when the radius of curvature approaches the thickness of a monolayer. Although slow relaxation beyond microseconds is observed in different perturbative simulations, the key structural features (e.g., dimension and shape of the fusion pore near the pore center) are consistent among independent simulations. These observations provide solid support for the use of coarse-grained and continuum models in the analysis of membrane remodeling. The combined coarse-grained and continuum analysis confirms the recent prediction of continuum models that the fusion pore is a metastable structure and that its optimal shape is neither toroidal nor catenoidal. Moreover, our results help reveal a new, to our knowledge, bowing feature in which the bilayers close to the pore axis separate more from one another than those at greater distances from the pore axis; bowing helps reduce the curvature and therefore stabilizes the fusion pore structure. The spread of the bilayer deformations over distances of hundreds of nanometers and the substantial reduction in energy of fusion pore formation provided by this spread indicate that membrane fusion can be enhanced by allowing a larger area of membrane to participate and be deformed.


[The role of arms color Doppler before creation arteriovenous fistula for hemodialysis].
our experience with color Doppler of arms blood vessels before creation arteriovenous fistula for hemodialysis. three years ago all patients who need surgery creation of arteriovenous fistula had Doppler ultrasound preoperative vascular mapping. Purpose of the view is to take a picture of proximal and distal part of arteria radials both arms, distal part of venous cephalica and proximal part of venous mediana antebrachii. We must know if these blood vessels have good morphological and hemodinamic criteria for creation arteiovenous fistula. in the study were twenty patients, creation of AV fistula was unsuccessful by two patients, but ultrasonic doctor had warning on very gracile blood vessels; by 18 patients in period of 2 to 36 months (mean 12.2 moths) we have optimal effective flow through dialysis filter 300 ml/min, and they are still in good function. carefully blood vessels review with color Doppler before creation arteriovenous fistula for hemodialysis, and marking suitable blood vessels with marker on the skin under sonography control reduced unsuccessful number of surgery exploration and destruction of blood vessels if AV fistula don't work. This is very important for hemodialysis patients, especially for diabetics.


The endoscopic endonasal approach to repair of iatrogenic and noniatrogenic cerebrospinal fluid leaks and encephaloceles of the anterior cranial fossa.
The current approach for the diagnosis and repair of spontaneous and traumatic anterior skull-base defects is oulined, highlighting the controversies that exist in the field and describing the strategies required to access different segments of the anterior cranial fossa. We reviewed the literature concerning endoscopic management of anterior skull-base defects. These publications have been combined with our own experience repairing cerebrospinal fluid (CSF) leaks and encephaloceles that developed spontaneously, traumatically, or intentionally as a result of endoscopic skull-base surgery. We present a systematic methodology for the repair of these defects. We have divided our surgical approach into four separate corridors. These are the transnasal, transsphenoidal, transethmoidal, and transmaxillary corridors. Dissection strategies vary for each corridor, but with a combination of approaches, all areas of the anterior skull base can be accessed. Skull-base defects are successfully repaired with a multilayered closure that often involves use of a vascularized pedicled mucosal flap. Adoption of this technique has decreased our rate of postoperative CSF leak from 5.9%-3.1%. Endoscopic endonasal repair of CSF leaks and encephaloceles has evolved significantly during the past decade. The versatility of different endoscopic approaches through the four endonasal corridors allows for the endoscopic repair of almost all skull-base defects. The use of vascularized pedicled mucosal flaps has evolved to cover these defects as part of multilayered closure strategies.


Urodynamic findings in female patients reporting with lower urinary tract symptoms.
The study has been undertaken to determine the reproducibility of lower urinary tract symptoms in female patients after urodynamic investigations. A Descriptive study with retrospective analysis of data. Combined Military Hospital Rawalpindi and Armed Forces Institute of Urology, Rawalpindi. Fifty consecutive women with a primary history of urinary incontinence were interviewed on a standard urinary incontinence questionnaire. They underwent structured clinical pelvic examination, along with cough stress test. Post void residual urine was measured. This was followed by a urine routine examination to exclude any urinary infection. Later these patients underwent a combination of urodynamic tests to predict their final diagnosis. The tests performed were filling cystometry and leak point pressures. The clinical diagnosis was found to have a variable reproducibility on urodynamic investigations. Clinical stress incontinence translated as genuine stress incontinence in 61.5% of the cases. Patients with isolated symptoms of stress incontinence had an incidence of detrusor instability up to 33.3%. For the symptoms of urgency and urge incontinence, the diagnosis was reproducible in up to 50% of the cases. In cases of mixed symptoms, 20% patients were found to have stress incontinence and 33.3% cases were found to have an unstable bladder, whereas in 46.6% of the cases no objective abnormality was found on urodynamic investigations. Urodynamic investigations should be performed in female patients with lower urinary tract symptoms, especially if irreversible procedure, e.g., surgery is being contemplated.


Plastidial Phosphoglucose Isomerase Is an Important Determinant of Seed Yield through Its Involvement in Gibberellin-Mediated Reproductive Development and Storage Reserve Biosynthesis in Arabidopsis.
The plastid-localized phosphoglucose isomerase isoform PGI1 is an important determinant of growth in Arabidopsis thaliana, likely due to its involvement in the biosynthesis of plastidial isoprenoid-derived hormones. Here, we investigated whether PGI1 also influences seed yields. PGI1 is strongly expressed in maturing seed embryos and vascular tissues. PGI1-null pgi1-2 plants had ∼60% lower seed yields than wild-type plants, with reduced numbers of inflorescences and thus fewer siliques and seeds per plant. These traits were associated with low bioactive gibberellin (GA) contents. Accordingly, wild-type phenotypes were restored by exogenous GA application. pgi1-2 seeds were lighter and accumulated ∼50% less fatty acids (FAs) and ∼35% less protein than wild-type seeds. Seeds of cytokinin-deficient plants overexpressing CYTOKININ OXIDASE/DEHYDROGENASE1 (35S:AtCKX1) and GA-deficient ga20ox1 ga20ox2 mutants did not accumulate low levels of FAs, and exogenous application of the cytokinin 6-benzylaminopurine and GAs did not rescue the reduced weight and FA content of pgi1-2 seeds. Seeds from reciprocal crosses between pgi1-2 and wild-type plants accumulated wild-type levels of FAs and proteins. Therefore, PGI1 is an important determinant of Arabidopsis seed yield due to its involvement in two processes: GA-mediated reproductive development and the metabolic conversion of plastidial glucose-6-phosphate to storage reserves in the embryo.


User perceptions on coastal resource state and management options in Curaçao.
Public environmental awareness and support for management measures are key determinants of the scope for successful implementation of natural resource management. To assess user perceptions and opinions on resource state and potential management options for the coastal zone of Curaçao, we queried 250 coastal resource users from around the island (sport divers, part-time artisanal fishermen and recreational boaters) using questionnaires. There is wide awareness of a long-term decline in coastal resource condition as measured by various indicators. Even among fishermen there was wide awareness of anthropogenic contributors to the decline, broad agreement of management measures required and a general willingness to contribute to management by means of annual license fees. Some of the more salient findings include the endorsement by fishermen of the current ban on spearfishing (81%), the regulation of the beach seine fishery for scad (77%), the introduction of fish reserves (72%), special protection for sea turtles (90%), conch (82%), and lobsters (72%), and notable support for gradual elimination of trap fisheries (45%). Also, both divers (65%) and boaters (92%) expressed the importance to them of an attractive coastline, with both groups expressing preference for natural (un-built) coastline (>74%) above other categories. Management based on the concept "user pays", as already implemented in the Netherlands Antilles on Saba and Bonaire, is well supported by the resource user public. A review of other main constraints such as finances and institutional capacity, shows that conditions are quite favorable for implementation of new legislation. Modem coral reef management is urgently needed in Curaçao to safeguard a key natural resource and concerted action is called for on the part of government agencies, legislators and elected officials.


MicroRNA-139-5p inhibits bladder cancer proliferation and self-renewal by targeting the Bmi1 oncogene.
MiR-139-5p has been reported to be overexpressed in many types of cancers, but its role in bladder cancer has not been elucidated yet. Here, we report that miR-139-5p functions as a tumor suppressor in bladder cancer and inhibits the cancer stem cell self-renewal by targeting Bmi1 directly. We found that miR-139-5p expression was significantly downregulated in the bladder cancer specimens compared with that in adjacent normal tissues. In vitro, restoration of miR-139-5p expression significantly inhibited the proliferation of bladder cancer cells. Mechanism analysis revealed that miR-139-5p could decrease Bmi1 protein levels by binding to the 3' untranslated region of Bmi1 messenger RNA. Stem cell-related proteins such as c-MYC, NANOG, OCT4, and KLF4 and signaling pathways such as Wnt signaling were suppressed by restoration of miR-139-5p in bladder cancer cells. In addition, miR-139-5p expression also blocked self-renewal of bladder cancer stem cells by inhibiting Bmi1. In summary, our study supports that miR-139-5p acts as a tumor suppressor in bladder cancer development and suppresses cancer stem cell property of bladder cancer. Our study also suggests that miR-139-5p has the potential to be used as a therapeutic molecule for bladder cancer treatment.


The effect of capsule-filling machine vibrations on average fill weight.
The aim of this paper is to study the effect of the speed of capsule filling and the inherent machine vibrations on fill weight for a dosator-nozzle machine. The results show that increasing speed of capsule filling amplifies the vibration intensity (as measured by Laser Doppler vibrometer) of the machine frame, which leads to powder densification. The mass of the powder (fill weight) collected via the nozzle is significantly larger at a higher capsule filling speed. Therefore, there is a correlation between powder densification under more intense vibrations and larger fill weights. Quality-by Design of powder based products should evaluate the effect of environmental vibrations on material attributes, which in turn may affect product quality.


Recommended standards for epidemiological surveillance systems for contagious bovine pleuropneumonia.
The initial draft of this document was prepared in June 1993 by the Ad hoc Group on Contagious Bovine Pleuropneumonia Surveillance Systems of the Office International des Epizooties. That draft was then amended in February 1997 by the Group of experts convened at the request of the International Committee to examine issues relating to the surveillance of the disease, with particular reference to the effects of vaccination programmes on surveillance systems. This updated report includes the following: epidemiological and other factors which influence the choice of surveillance systems. Sampling and surveillance strategies. Diagnostic methods applicable to surveillance systems. The repercussions of vaccination on surveillance systems.


Dopaminergic neurones in the zona incerta exert a stimulatory control on gonadotrophin release via D1 dopamine receptors.
The zona incerta (ZI) is a site of dopamine nerve terminals and part of the incertohypothalamic tract (I-H). Previous findings indicate that dopamine in the ZI has a stimulatory control on the release of luteinizing hormone (LH) and occurrence of ovulation. The effect of acute administration into anaesthetised rats of selective D1 and D2 dopamine agonists and antagonists injected into the ZI on plasma luteinizing hormone (LH) and on the occurrence of ovulation has now been investigated. It was found that bilateral injections on the day of pro-oestrus of a selective D1 antagonist, Sch 23390, inhibited ovulation at 10 micrograms/side/rat. Unilateral injections of a selective D1 agonist, SKF 38393, at 10 micrograms/rat stimulated a significant rise in plasma LH concentration in ovariectomised oestrogen-primed rats, and this was partially reversed by systemic pre-treatment with Sch 23390. The selective D2 agonist, LY 171555, and D2 antagonists, sulpiride and domperidone, had no effect on plasma LH levels or ovulation. This indicates that D1 receptors (but not D2 receptors) in the ZI are involved in the control of gonadotrophin release and may have a physiological function in reproductive processes.


Integrative analysis of metabolome and gut microbiota in Patients with pancreatic ductal adenocarcinoma.
Pancreatic ductal adenocarcinoma (PDAC) is a highly malignant tumour with a poor prognosis and a high mortality rate. It is of great significance to explore sensitive or specific biomarkers for early diagnosis and prognosis. We first examined the metabolome and gut microbiota of resectable and unresectable PDAC patients to comprehensively investigate the characteristics of PDAC at different stages of progression. At the genus level, we found that the relative abundances of Alistipes, Anaerostipes, Faecalibacterium and Parvimonas were reduced in unresectable PDAC patients, whereas Pseudonocardia, Cloacibacterium, Mucispirillum, and Anaerotruncus were increased. Metabolomics analysis showed that the main changed metabolites were amino acids, carnitine derivatives, lipids and fatty acids. ROC analysis showed that Oleic acid, Linoleic acid, Palmitic acid, Linoelaidyl carnitine, 2-Octenedioic acid, 3R, 7R-1,3,7-Octanetriol, LysoPE (P-16:0/0:0) and 3-Hydroxyanthranilic acid had high AUC values (>0.9). Function and network analyses showed that these altered metabolites correlated with NF-kappa B signalling, the FXR/RXR pathway, mitochondrial dysfunction, mTOR signalling and IL-6 signalling. In particular, the abundance of Palmitic acid, Oleic acid, Linoelaidyl carnitine and 2-Octenedioic acid positively correlated with g_Anaerostipes, g_Alistipes, s_indistinctus, s_catus and s_formicigenerans but negatively correlated with g_Cloacibacterium, s_reuteri and s_hathewayi. Meanwhile,We also found that s_catus, s_ formicigenerans, s_ hathewayi, g_ Alistipes, g_ Anaerostipes, PE (22:6 (4Z, 7z, 10z, 13z, 16Z, 19Z)/p-18:1 (11z)), (3R, 7R) - 1,3,7-octanetriol and linoelaidyl carnitine were positively correlated with the survival time of patients.These findings may be helpful for the differentiation of resectable and unresectable PDAC based on changes in intestinal flora and metabolites at different stages of PDAC. This study also provides a strategy for preventing the deterioration of PDAC by regulating the gut microbiota and metabolism.


Relaxin Attenuates Contrast-Induced Human Proximal Tubular Epithelial Cell Apoptosis by Activation of the PI3K/Akt Signaling Pathway In Vitro.
Background. Contrast-induced acute kidney injury (CI-AKI) is one of the main causes of iatrogenic acute kidney injury (AKI); however, therapeutic strategies for AKI remain limited. This study aims to explore the effect of relaxin (RLX) on contrast-induced HK-2 apoptosis and its underlying mechanisms. Methods. Renal tubular epithelial cells (HK-2) were incubated either with or without ioversol, human H2 relaxin, and LY294002 (the inhibitor of the PI3K/Akt signal pathway). Cell viability was evaluated with a CCK-8 assay. Apoptotic morphologic alterations were observed using the Hoechst 33342 staining method. Apoptosis was detected with Annexin V staining. Western blot analysis was employed to measure the expression of pAkt (S473), Akt, cleaved caspase-3, Bcl-2, Bax, and actin proteins. Results. Ioversol reduced the viability of HK-2 cells. Western blotting results revealed decreased expression of phosphorylated Akt in cells treated with ioversol. The activities of caspase-3 and Bax protein increased, while the expression of Bcl-2 protein decreased. As a result, the Bax/Bcl-2 ratio increased after treatment with ioversol. These effects were reversed when HK-2 cells were cotreated with RLX. However, with preadministration of PI3K/Akt pathway inhibitor LY294002, the effect of RLX was blocked. Conclusion. Our study demonstrates that relaxin attenuates ioversol induced cell apoptosis via activation of the PI3K/Akt signaling pathway, suggesting that RLX might play a protective role in the treatment of CI-AKI.


Tissue engineering: in vitro embryonal nidation in a murine endometrial construct.
The epithelial and mesothelial cellular components of organs can be obtained as dissociated cells using adequate procedures of enzymatic digestion followed by pycnotic separation on density gradients. Using a specially developed procedure for tissue dissociation, the epithelial and connective tissue components of endometria from pseudopregnant mice were grown in culture using a combination of three dimensional culture of connective tissue components in collagen gel, with the superimposition of epithelial components in liquid medium on the surface of the gells. After a few days of growth, when the cultures became dense, murine blastocysts obtained on postcoital day 4.5 by fallopian flushing of hormonally primed and mated mice, were transferred onto the imitated endometria. The blastocysts hatched and grew on the endometrial epithelium as spherical coherent conglomerates of cells quite different from hatched blastocysts grown on the surface of a petri dish, in which the presumtive trophoblasts spred around the central mass. Light and electronmicroscopy of resin embeded sections (2 days after nidation on the simulated endometria) revealed that at least two populations of cell types were recognisable as layers. This is interpreted as an early sign of morphogenesis and the first visible steps of differentiation. The presence of mitotic figures indicates viability and continuing growth. Electronmicroscopy of cell types grown under conditions simulating in vivo tissue architectonics showed overtly less cytopathology and better cell function. Simulated endometria may, therefore, serve as an attractive model for studying early mammalian embryogenesis and the effects of toxic agents.


One lung ventilation in a patient with an upper and lower airway abnormality.
One-lung ventilation for a thoracotomy procedure was achieved with the help of a endobronchial blocker in a young girl with limited mouth opening, minimal neck extension, and a distorted tracheo-bronchial anatomy. As the patient would not cooperate for an awake nasotracheal intubation despite adequate preperation, an inhalational anesthetic was used to make the patient unconscious, taking care that spontaneous breathing was maintained. Nasotracheal intubation was done with the help of a fiberoptic bronchoscope. A wire-guided Arndt endobronchial blocker was placed coaxially through the endotracheal tube using a fiberoptic bronchoscope. This case report highlights that in a scenario of both upper and lower airway distortion, a bronchial blocker positioned through a nasotracheal tube under fiberoptic guidance might be the best option when one-lung ventilation is required.


Measuring adherence to medication in schizophrenia: the relationship between attitudes toward drug therapy and plasma levels of new-generation antipsychotics.
Nonadherence to medication is still a major problem in the treatment of schizophrenia. The current longitudinal study investigated whether the patients' attitudes toward treatment correlated with the ratio of observed vs expected plasma levels of antipsychotic drugs as an objective measurement of adherence. Data of patients starting monotherapy with a new-generation antipsychotic were collected 2, 4, and 12 weeks after the initiation of treatment. Next to the assessment of patients' attitudes toward medication by means of the Drug Attitude Inventory, the ratio of the observed vs expected plasma level was calculated. Antipsychotic-induced side effects were evaluated by means of the Udvalg for Kliniske Undersogelser Side Effect Rating Scale. A total of 93 patients were eligible for statistical analysis. About one-half of the ratios of observed vs expected plasma levels ranged from 0.5 to 2 and were considered normal, whereas the other ratios were considered either too low (<0.5) or too high (>2). No consistent correlation between patients' attitude toward drug therapy and the individual ratios of observed vs expected plasma levels of medication was detected. This finding was not affected by side effects. Our results highlight the importance of recognizing the complex nature of adherence to medication in schizophrenia patients. Importantly, we found no consistent correlation between subjective and objective measures of medication adherence. Therefore, monitoring adherence to medication remains a challenge in clinical practice.


Induction of labour using balloon catheter and prostaglandin gel.
The process of induction of labour (IOL) commonly involves cervical ripening, using mechanical or pharmacological methods. However, the optimum protocol of IOL remains undetermined. To report the maternal, neonatal and process outcomes for women being induced with a balloon catheter or prostaglandin E2 gel (PGE2). Matched retrospective cohort study of women requiring IOL at term with a modified Bishop's score (MBS) <7. Those who underwent cervical ripening with a double balloon catheter were matched 1:1 for parity, indication and MBS score to those using PGE2 gel. The primary outcome measure was mode of birth. After exclusions, data from 427 women induced with a balloon catheter were able to be matched to 427 women induced with PGE2 gel. Compared to women receiving PGE2 gel, those induced using a balloon catheter were more likely to have an unassisted vaginal birth (50.4% vs 42.9%; P = 0.028), a lower mean estimated blood loss (420 vs 481 mL; P = 0.028), a reduction in fetal acidaemia (2.4% vs 8.8%; P = 0.003) and a greater likelihood experiencing vaginal delivery within 24 h (33.7% vs 25.8%; P = 0.011). Similar to findings in randomised controlled trials and meta-analyses of IOL methods, we observed more unassisted vaginal births, less blood loss, decreased fetal acidaemia, and a greater likelihood of vaginal delivery within 24 h, with use of balloon catheter as compared to PGE2 vaginal gel.


Biological and Pharmacological Aspects of the NK1-Receptor.
The neurokinin 1 receptor (NK-1R) is the main receptor for the tachykinin family of peptides. Substance P (SP) is the major mammalian ligand and the one with the highest affinity. SP is associated with multiple processes: hematopoiesis, wound healing, microvasculature permeability, neurogenic inflammation, leukocyte trafficking, and cell survival. It is also considered a mitogen, and it has been associated with tumorigenesis and metastasis. Tachykinins and their receptors are widely expressed in various human systems such as the nervous, cardiovascular, genitourinary, and immune system. Particularly, NK-1R is found in the nervous system and in peripheral tissues and are involved in cellular responses such as pain transmission, endocrine and paracrine secretion, vasodilation, and modulation of cell proliferation. It also acts as a neuromodulator contributing to brain homeostasis and to sensory neuronal transmission associated with depression, stress, anxiety, and emesis. NK-1R and SP are present in brain regions involved in the vomiting reflex (the nucleus tractus solitarius and the area postrema). This anatomical localization has led to the successful clinical development of antagonists against NK-1R in the treatment of chemotherapy-induced nausea and vomiting (CINV). The first of these antagonists, aprepitant (oral administration) and fosaprepitant (intravenous administration), are prescribed for high and moderate emesis.


Etiology of solitary pulmonary nodules in patients with human immunodeficiency virus infection.
We assessed 10 patients with human immunodeficiency virus (HIV) infection (nine of whom had AIDS) and solitary pulmonary nodules (SPNs) that were detected on roentgenograms. Five of the patients presented with respiratory symptoms. The etiology of the SPN was determined for eight of these patients: six had infections (hydatidosis, mucormycosis, or infection with Nocardia asteroides, Cryptococcus neoformans, cytomegalovirus, or Pneumocystis carinii), one had non-Hodgkin's lymphoma, and the remaining patient had round atelectasis. Sputum studies were of no diagnostic value in any of these cases. Fiberoptic bronchoscopy with bronchoalveolar lavage and transbronchial biopsy was diagnostic in two of seven cases, and percutaneous transthoracic needle biopsy (PTNB) was diagnostic in one of four cases. Several microorganisms that were not the cause of the SPNs were observed in samples of sputum, bronchoscopic specimens, and PTNB specimens. Thoracotomy was diagnostic in the three cases in which it was performed. We conclude that the management of SPNs in HIV-infected patients is complicated by the low sensitivity and specificity of the diagnostic tests used.


Nrf2 is essential for timely M phase entry of replicating hepatocytes during liver regeneration.
The transcription factor nuclear factor erythroid 2-related factor 2 (Nrf2) regulates various cellular activities, including redox balance, detoxification, metabolism, autophagy, proliferation, and apoptosis. Several studies have demonstrated that Nrf2 regulates hepatocyte proliferation during liver regeneration. The aim of this study was to investigate how Nrf2 modulates the cell cycle of replicating hepatocytes in regenerating livers. Wild-type and Nrf2 null mice were subjected to 2/3 partial hepatectomy (PH) and killed at multiple time points for various analyses. Nrf2 null mice exhibited delayed liver regrowth, although the lost liver mass was eventually restored 7 days after PH. Nrf2 deficiency did not affect the number of hepatocytes entering the cell cycle but did delay hepatocyte mitosis. Mechanistically, the lack of Nrf2 resulted in increased mRNA and protein levels of hepatic cyclin A2 when the remaining hepatocytes were replicating in response to PH. Moreover, Nrf2 deficiency in regenerating livers caused dysregulation of Wee1, Cdc2, and cyclin B1 mRNA and protein expression, leading to decreased Cdc2 activity. Thus, Nrf2 is required for timely M phase entry of replicating hepatocytes by ensuring proper regulation of cyclin A2 and the Wee1/Cdc2/cyclin B1 pathway during liver regeneration.


Enzymatic synthesis of fructose 1,6-diphosphate with ATP regeneration in a batch reactor and a semibatch reactor using purified enzymes of Bacillus stearothermophilus.
The enzymatic synthesis of fructose 1,6-diphosphate (FDP), an important glycolytic intermediate whose applications in the field of medicine have generated a great deal of interest, was performed in a batch reactor and a semibatch reactor. Using the batch reactor, FDP was first synthesized from glucose by three enzymatic reactions and the ATP consumed was regenerated simultaneously using conjugated enzymes, all of which were purified from crude cell extract of thermophilic Bacillus stearothermophilus. The results of the experiments performed using several enzyme concentrations suggest the existence of an optimum concentration for each enzyme at which the maximum FDP yield can be attained. Since the thermal decomposition of acetyl phosphate reduced the yield of FDP in the batch reactor, the use of a semibatch reactor in which acetyl phosphate was fed continuously was examined. The yield of FDP was improved but the time required to complete the reaction was longer, resulting in a lower productivity of FDP. The yields observed in the two reactors using various enzyme and substrate concentrations were in good agreement with the theoretical predictions calculated based on differential equations derived for the system using the rate equations and the kinetic parameters determined previously. This means that these equations can be used for the analysis of the experimental results as well as for determining the optimum experimental conditions.


A randomized-controlled trial comparing supervised aerobic training to resistance training followed by unsupervised exercise on physical functioning in older breast cancer survivors.
This study compared the relative efficacy of aerobic training to resistance training on physical functioning in older breast cancer survivors and determined whether benefits could be maintained by transitioning to unsupervised home-based training. Early-stage, post-treatment, older (≥65 years) breast cancer survivors (n = 114; mean age 72 years) were randomized to 12 months of supervised aerobic (n = 37), resistance (n = 39) or stretching (active control; n = 38) training followed by 6 months of unsupervised home-based training. Outcomes included aerobic capacity by 6-min walk distance (6MWD; m), maximal upper and lower body strength (1-repetition maximum; kg); physical function by short physical performance battery (SPPB), SF-36 and Late Life Function and Disability Instruments. Over 12-months of supervised exercise, all groups improved in muscle strength and SPPB scores, but resistance trained women also improved 6MWD. Improvements in upper and lower body strength in the resistance group were significantly greater than those in the stretching control (+2.5 kg vs. +1.8 kg; p = 0.05) and aerobic groups (+8.3 kg vs +2.7 kg; p = 0.047), respectively, with trends for greater improvements in 6MWD (+57.9 m vs. +22.5 m; p = 0.057) and self-report physical function (+4.8 vs. -4.4; 0.066) in resistance trained women versus controls. Compared to values at 12 months, there were no changes during unsupervised training in any measure within or between groups, except for self-reported advanced lower extremity function which improved in the resistance group and fell in the aerobic group (+1.3 vs. -3.1; p = 0.043). Supervised exercise can improve strength and physical functioning among older breast cancer survivors. Resistance training may lead to better improvements compared to aerobic or flexibility training, whether in a supervised or unsupervised setting. Clinicaltrials.govNCT00662103.


On-farm mortality, causes and risk factors in Estonian beef cow-calf herds.
High on-farm mortality is associated with lower financial return of production and poor animal health and welfare. Understanding the reasons for on-farm mortality and related risk factors allows focus on specific prevention measures. This retrospective cohort study used cattle registry data from the years 2013 and 2014, collected from cattle from all Estonian cow-calf beef herds. The dataset contained 78,605 animal records from 1321 farms in total. Including unassisted deaths and euthanasia (2199 in total) the on-farm mortality rate was 2.14 per 100 animal-years. Across all age groups of both sexes the mortality rate (MR) was highest for bull calves up to three months old (MR=7.78 per 100 animal-years, 95% CI 6.97; 8.68) followed by that for heifer calves (MR=6.21 per 100 animal-years, 95% CI 5.49; 7.02). For female cattle the mortality risk declined after three months of age but increased again among animals over 18 months. The reason for death stated by the farmers was analysed for cattle under animal performance testing. Other/unknown reasons, trauma and accidents, as well as metabolic and digestive disorders, formed the three most commonly reported reasons for death in cattle of all age groups. Weibull proportional hazard models with farm frailty effects were applied in three age categories (calves up to three months, youngstock from three to 18 months and cattle aged over 18 months) to identify factors associated with the risk of mortality. Male sex was associated with increased risk of mortality in cattle up to 18 months of age. No difference between breeds was found for cattle up to 18 months of age. Beef cattle breeds rarely represented or dairy breeds (breed category 'Other') had the highest mortality hazard (HR=1.41, 95% CI 1.11; 1.78) compared to Hereford. The hazard of mortality generally increased with herd size for calves, young stock and older bulls. In female cattle over 18 months of age there was no difference in mortality hazard over herd size categories. Herd location was controlled in the models and regional differences in mortality hazard were found. Common to all age groups, calving season was associated with increased risk of mortality.


What causes the word gap? Financial concerns may systematically suppress child-directed speech.
Parents with fewer educational and economic resources (low socioeconomic-status, SES) tend to speak less to their children, with consequences for children's later life outcomes. Despite this well-established and highly popularized link, less research addresses why the SES "word gap" exists. Moreover, while research has assessed individual-level contributors to the word gap-like differences in parenting knowledge-we know little about how structural constraints that vary according to SES might affect caregivers' speech. In two pre-registered studies, we test whether experiencing financial scarcity can suppress caregivers' speech to their children. Study 1 suggests that higher-SES caregivers who are prompted to reflect on scarcity-particularly those who reflect on financial scarcity-speak less to their 3-year-olds in a subsequent play session, relative to a control group. Study 2 suggests that mid- to higher-SES caregivers engage in fewer back-and-forth exchanges with their children at the end of the month-when they are more likely to be experiencing financial hardship-than the rest of the month. These studies provide preliminary evidence that-above and beyond caregivers' individual characteristics-structural constraints may affect how much parents speak to their children.


Impact of vegan diets on gut microbiota: An update on the clinical implications.
Numerous studies indicate that microbiota plays an important role in human health. Diet is a factor related to microbiota which also influences human health. The relationships between diet, microbiota, and human health are complex. This review focuses on the current literature on vegan diets and their unique impact on gut microbiota. We also report on the health benefits of a vegan diet for metabolic syndrome, cardiovascular disease, and rheumatoid arthritis concerning relevant impacts from gut microbiota. Despite evidence supporting the clinical relevance of vegan gut microbiota to human health, the whole mechanism awaits further investigation.


[Ergonomic aspects in the prevention of occupational cumulative trauma disorders of the locomotor system].
In recent years, a rapid increase in the number of cumulative trauma disorders (CTDs) induced by repeatability of certain functions in the course of work performance has been observed. In the United States, a twenty-fold increase in the number of registered CTDs of upper limbs has been noted during the last ten years. For example in 1993, CTDs constituted 60% of all occupational diseases. A similar observation was made in Australia, Canada, Japan, Sweden, the United Kingdom and other European countries. It is thought that this high incidence of cumulative trauma disorders result, to a great extent, from changes in technologies leading finally to almost unilateral involvement of the human locomotor system. Ergonomics plays a key role in the prevention of the locomotor system occupational traumas, since as the only one discipline, it is able to provide criteria for securing safety work without disturbing its inherent sense, namely the efficiency. Risk factors reported in Western countries will also emerge in Polish economy in the near future. In Poland like in other countries, employers, managers of work posts, safety and hygiene services, as well as occupational physicians frequently fail to identify CTD risk factors which occur in the working environment. In this paper, the author presents the literature data on CTD etiology, and the criteria for identifying occupational risk factors. The literature review clearly indicates that the studies of cumulative trauma disorders of the musculoskeletal system has recently been intensified. But the evaluation criteria, nomenclature and analytical methods still needs to be standardized.


Suspected orbital myositis associated with COVID-19.
A 44-year-old male patient developed proptosis, edema, and erythema progressing to complete ptosis and supraduction deficit 2 days after positive COVID-19 test. He failed to improve on systemic antibiotics. MRI showed thickening and T2 enhancement of the superior rectus/levator complex consistent with orbital myositis. He improved on intravenous corticosteroids and experienced continued gradual improvement on oral steroids.


Surgical Health Needs of Minor Refugees in Germany: A Cross-Sectional Study.
There has been a substantial rise in refugees entering Germany over the past years, of which approximately one-third are underaged. Many end up in pediatric surgical care, and little is known about the health of these individuals. Our study was designed to assess the surgical-related health status of underage refugees based on a large sample cohort. After ethics board approval, we used a structured questionnaire to collect demographic information and surgical health-related elements in three large refugee accommodation centers. A total of 461 minor refugees were included. The majority were boys (54.5%) with an average age of 8 years. Out of the eight recorded countries of origin, most children came from Syria (33.6%) followed by Afghanistan (23.2%). Previous operative interventions were recorded in 42.2% of participants. Among girls, 11% suffered genital mutilation. Trauma was common and the most common mechanism was a fall from bicycle (38%) followed by burn injuries (7.4%). Up to 20% of them experienced physical violence during the flight or in the accommodation facility. Vaccination rates varied widely according to origin. Of the participants, only 63% were vaccinated according to schedule. Chronic diseases were found in only 13% of the study cohort, anemia being most prevalent at 4%. Minor refugees have specific health-related problems that must be considered to ensure appropriate medical care. Many refugee children were victims of physical violence and many girls suffered genital mutilation. Vaccination status is unreliable; therefore, tetanus vaccination should always be considered when these patients seek pediatric surgical care. Tailored anticipatory guidance should be provided to this patient population.


Assessment of right ventricular infarction with contrast-enhanced magnetic resonance imaging.
Evaluation of contrast-enhanced magnetic resonance imaging to assess right ventricular infarction in patients with acute inferior myocardial infarction. Contrast-enhanced magnetic resonance imaging has been used for assessing scar tissue after left ventricular infarction. The value of contrast-enhanced magnetic resonance imaging to assess right ventricular infarction is unknown and was evaluated. Consecutive patients (n=18) with first acute inferior infarction were included. Resting electrocardiogram and right-sided electrocardiogram were acquired to assess right ventricular involvement. Resting cine magnetic resonance imaging was performed to evaluate right ventricular function and volumes, whereas the extent of right ventricular scar tissue was assessed by contrast-enhanced magnetic resonance imaging. Cine magnetic resonance imaging was repeated at 6-months follow-up to re-assess right ventricular function and volumes. Sensitivity and specificity of magnetic resonance imaging were 100 and 78%, respectively, to detect right ventricular infarction (using the right-sided electrocardiogram as the gold standard). At 6 months follow-up, patients with scar tissue on contrast-enhanced magnetic resonance imaging showed right ventricular dilatation. Moreover, the extent of right ventricular scar tissue was linearly related to the severity of right ventricular dilatation. Contrast-enhanced magnetic resonance imaging permits accurate assessment of right ventricular scar tissue. Patients with extensive right ventricular infarction demonstrate right ventricular dilatation at 6 months follow-up.


Single-cell analyses of X Chromosome inactivation dynamics and pluripotency during differentiation.
Pluripotency, differentiation, and X Chromosome inactivation (XCI) are key aspects of embryonic development. However, the underlying relationship and mechanisms among these processes remain unclear. Here, we systematically dissected these features along developmental progression using mouse embryonic stem cells (mESCs) and single-cell RNA sequencing with allelic resolution. We found that mESCs grown in a ground state 2i condition displayed transcriptomic profiles diffused from preimplantation mouse embryonic cells, whereas EpiStem cells closely resembled the post-implantation epiblast. Sex-related gene expression varied greatly across distinct developmental states. We also identified novel markers that were highly enriched in each developmental state. Moreover, we revealed that several novel pathways, including PluriNetWork and Focal Adhesion, were responsible for the delayed progression of female EpiStem cells. Importantly, we "digitalized" XCI progression using allelic expression of active and inactive X Chromosomes and surprisingly found that XCI states exhibited profound variability in each developmental state, including the 2i condition. XCI progression was not tightly synchronized with loss of pluripotency and increase of differentiation at the single-cell level, although these processes were globally correlated. In addition, highly expressed genes, including core pluripotency factors, were in general biallelically expressed. Taken together, our study sheds light on the dynamics of XCI progression and the asynchronicity between pluripotency, differentiation, and XCI.


Excitatory muscarinic modulation strengthens virtual nicotinic synapses on sympathetic neurons and thereby enhances synaptic gain.
Acetylcholine excites many neuronal types by binding to postsynaptic m1-muscarinic receptors that signal to ion channels through the G(q/11) protein. To investigate the functional significance of this metabotropic pathway in sympathetic ganglia, we studied how muscarinic excitation modulated the integration of virtual nicotinic excitatory postsynaptic potentials (EPSPs) created in dissociated bullfrog B-type sympathetic neurons with the dynamic-clamp technique. Muscarine (1 muM) strengthened the impact of virtual synapses by reducing the artificial nicotinic conductance required to reach the postsynaptic firing threshold from 20.9 +/- 5.4 to 13.1 +/- 3.1 nS. Consequently, postganglionic action potential output increased by 4-215% when driven by different patterns of virtual presynaptic activity that were chosen to reflect the range of physiological firing rates and convergence levels seen in amphibian and mammalian sympathetic ganglia. In addition to inhibiting the M-type K(+) conductance, muscarine activated a leak conductance in three of 37 cells. When this leak conductance was reproduced with the dynamic clamp, it also acted to strengthen virtual nicotinic synapses and enhance postganglionic spike output. Combining pharmacological M-conductance suppression with virtual leak activation, at resting potentials between -50 and -55 mV, produced synergistic strengthening of nicotinic synapses and an increase in the integrated postganglionic spike output. Together, these results reveal how muscarinic activation of a branched metabotropic pathway can enhance integration of fast EPSPs by modulating their effective strength. The results also support the hypothesis that muscarinic synapses permit faster and more accurate feedback control of autonomic behaviors by generating gain through synaptic amplification in sympathetic ganglia.


[Laparoscopic total mesorectal excision for extraperitoneal rectal cancer. Oncological outcome at 5 years].
Total mesorectal excision (TME) is the cornerstone of surgical treatment for extraperitoneal rectal cancer. The aim of the present study was to analyse our five-year experience with laparoscopic TME, evaluating the overall five-year and disease-free survival rates. Twenty-five patients with low-middle rectal cancer were treated with laparoscopic TME. Patients with advanced rectal cancer were treated preoperatively with neoadjuvant radiochemotherapy. Five-year overall survival and disease-free survival were calculated according to the Kaplan-Meier method. Twenty-three ultralow anterior resections with Knight-Griffen anastomosis and 3 abdominoperineal resections were performed. At 30 days mortality was zero, while morbidity was 20% (all minor complications). The mean follow-up period was 30.5 months. Five-year overall survival was 80.2%, and five-year disease-free survival 80.9%. Our experience shows that laparoscopic TME is a safe and oncologically correct procedure. Oncologic outcomes were comparable to those reported in all major international experiences, and the results were very similar to those obtained with the laparotomic approach. However, it remains a complex technique, requiring an adequate learning curve. More prospective, randomised trials are needed in order to define laparoscopic TME as the new gold standard for the treatment of extraperitoneal rectal cancer.


Incisional hernia following laparoscopy: a survey of the American Association of Gynecologic Laparoscopists.
To estimate the risk factors for and incidence of post-laparoscopy incisional hernia. A questionnaire was sent to all individuals on a mailing list from the American Association of Gynecologic Laparoscopists. Adequately completed responses were entered into a computer-based data bank for analysis. Of the 11,500 surveys mailed, 3293 were returned; of these, 3217 were evaluable. A total of 933 hernias was reported from an estimated 4,385,000 laparoscopic procedures (an incidence of 21 per 100,000); 167 (17.9%) were reported to have occurred despite fascial closure. Six hundred sixty-five patients (71:3%) had subsequent surgical repair. Seven hundred twenty-five (86.3%) of the 840 hernias in which the size of the original fascial defect was noted, occurred in sites where ports 10 mm in diameter or larger had been placed. The occurrence of hernias is a function of the number of laparoscopies performed (P < .0001) and is not related to the length of the surgeon's career (P = .41). In at least 157 instances (16.8%), the presenting symptom or morbidity of the hernia was directly related to the involvement of the large or small intestine. Post-laparoscopy incisional hernias occur at an approximate incidence of 21 per 100,000 and are associated with significant morbidity. These hernias are most likely to occur when large ports are used. As currently practiced, closure is not completely protective. Further methods or devices should be developed to minimize the risk of hernia formation.


The leptin-like effects of 3-d peripheral administration of a melanocortin agonist are more marked in genetically obese Zucker (fa/fa) than in lean rats.
The effects of a 3-d peripheral administration of an alpha-MSH agonist, MTII, on body weight and the expression of uncoupling proteins (UCPs) and carnitine palmitoyltransferase-1 were determined in lean and genetically obese fa/fa rats by comparing MTII-treated animals with two different control groups, one being ad libitum fed, the other pair-fed to the amount of food consumed by MTII-treated rats. MTII treatment of lean and obese rats lowered food intake and body weight, the effects being more marked in obese than in lean rats. In both groups, MTII administration suppressed the increased plasma FFA levels brought about by food restriction. In lean rats, MTII prevented the decrease in brown adipose tissue UCP1, UCP2, and UCP3 expression and muscle UCP3 occurring during food restriction. In obese animals, MTII markedly increased brown adipose tissue (7-fold) and muscle (2.5-fold) UCP3 expression. The decrease in liver carnitine palmitoyltransferase-1 elicited by food restriction in lean and obese rats was prevented by MTII administration. In summary, the effects of MTII resemble those of leptin and are more marked in obese than in lean animals, in keeping with their reported reduced endogenous melanocortin tone. Melanocortin agonists may be useful in the treatment of obesity associated with impaired leptin signaling.


Intrinsic ankle stiffness during standing increases with ankle torque and passive stretch of the Achilles tendon.
Individuals may stand with a range of ankle angles. Furthermore, shoes or floor surfaces may elevate or depress their heels. Here we ask how these situations impact ankle stiffness and balance. We performed two studies (each with 10 participants) in which the triceps surae, Achilles tendon and aponeurosis were stretched either passively, by rotating the support surface, or actively by leaning forward. Participants stood freely on footplates which could rotate around the ankle joint axis. Brief, small stiffness-measuring perturbations (<0.7 deg; 140 ms) were applied at intervals of 4-5 s. In study 1, participants stood at selected angles of forward lean. In study 2, normal standing was compared with passive dorsiflexion induced by 15 deg toes-up tilt of the support surface. Smaller perturbations produced higher stiffness estimates, but for all perturbation sizes stiffness increased with active torque or passive stretch. Sway was minimally affected by stretch or lean, suggesting that this did not underlie the alterations in stiffness. In quiet stance, maximum ankle stiffness is limited by the tendon. As tendon strain increases, it becomes stiffer, causing an increase in overall ankle stiffness, which would explain the effects of leaning. However, stiffness also increased considerably with passive stretch, despite a modest torque increase. We discuss possible explanations for this increase.


Senescent fibroblasts promote neoplastic transformation of partially transformed ovarian epithelial cells in a three-dimensional model of early stage ovarian cancer.
Most epithelial ovarian cancers are diagnosed postmenopausally, although the well-established epidemiological risk factors (parity, oral contraceptive use) are premenopausal. We hypothesized that accumulation of senescent fibroblasts, together with concomitant loss of presenescent fibroblasts within the ovarian cortex, promotes initiation and early development of ovarian cancer from ovarian surface epithelial (OSE) cells. To test this, we established immortalized OSE (IOSE) cell lines that mimic early neoplastic transformation by overexpressing the CMYC oncogene (IOSE(CMYC)) and normal ovarian presenescent (PSN) and senescent (SEN) fibroblast cell lines. We then evaluated the ability of PSN and SEN fibroblasts to transform IOSE and IOSE(CMYC) after coculture. SEN fibroblasts significantly enhanced neoplastic development of IOSE(CMYC) cells; there was an up to 15-fold increase in migration of IOSE(CMYC) cells cocultured with SEN fibroblasts compared with PSN fibroblasts. Conditioned medium from SEN fibroblasts promoted anchorage-independent growth of IOSE(CMYC) cells. We studied fibroblast-epithelial cell interactions in heterotypic three-dimensional spheroid models. Dual immunohistochemical staining of spheroids for a proliferation marker (MIB-1) and cytokeratin-18 indicated that SEN fibroblasts induce approximately a five-fold increase in proliferation of IOSE(CMYC) cells relative to cocultures with PSN fibroblasts. SEN, but not PSN fibroblasts, also induced nuclear atypia in epithelial cells in three-dimensional spheroids. These data suggest for the first time that the accumulation of senescent, or loss of presenescent fibroblasts, can promote neoplastic development of partially transformed OSE cells in vitro and illustrates the power of using three-dimensional heterotypic modeling to gain better insights into the etiology underlying the development of epithelial ovarian cancer.


Magnetic circular dichroism of porphyrins containing M = Ca, Ni, and Zn. A computational study based on time-dependent density functional theory.
A theoretical study is presented on the magnetic circular dichroism (MCD) exhibited by the porphyrin complexes MP (M = Mg,Ni,Zn), MTPP (M = Mg,Ni,Zn), and NiOEP, where P = porphyrin, TPP = tetraphenylporphyrin, and OEP = octaethylporphyrin. The study makes use of a newly implemented method for the calculation of A and B terms from the theory of MCD and is based on time-dependent density functional theory (TD-DFT). It is shown that the MCD spectrum is dominated by a single positive A term in the Q-band region in agreement with experiment where available. The band can be fully explained as the first transition in Gouterman's four-orbital model for the type of porphyrins studied here. For the Soret band, the experimental MCD spectrum appears as a single positive A term. This is also what is found computationally for NiP and NiTPP, where the second transition in Gouterman's four-orbital model give rise to a positive A term. However, for the remaining systems, the simulated MCD spectrum is actually due to two B terms that have the appearance of one positive pseudo A term. The two B terms appear because the second Gouterman state is coupled strongly to a second excited state (b(2u) --> 2e(g)) of nearly the same energy by the external magnetic field.


Prenatal Diagnosis of a Persistent Fifth Aortic Arch, Pulmonary-to-Systemic type: An Unusual Association with Evolving Aortic Coarctation.
Persistent fifth arch (PFA) is a rare anomaly that is often underdiagnosed and missed. Different PFA types that have been reported in the literature are systemic-to-systemic type (most common), systemic-to-pulmonary artery (PA), and PA-to-systemic types. The designations of systemic-to-PA or PA-to-systemic are based on if the PFA is a source of PA or systemic blood flow, respectively, in the setting of critical proximal obstruction (pulmonary atresia or aortic atresia). This case describes an unusual PFA, which connects the distal PA to distal ascending aorta; however, it is not associated with critical proximal obstruction, and while it appeared to be an incidental finding in early gestation, progressive serial distal obstruction of the left fourth arch was seen to develop. This case highlights that prenatal diagnosis of PFA is possible and that once a diagnosis is made, serial fetal echocardiograms should be performed to evaluate for evolving lesions.


Metastatic large cell lung cancer presenting with numb chin syndrome.
The numb chin syndrome is an uncommon occurrence with cancer and consists of numbness in the area of distribution of the mental nerve. Most cases of this syndrome occur in the presence of underlying breast cancer or lymphoma with survival measured in months once this syndrome is manifest. Reported below is the case of a man with metastatic large cell lung cancer whose presenting complaint was chin and lip numbness.


Access to Transportation and Health Care Visits for Medicaid Enrollees With Diabetes.
Diabetes is a chronic condition that requires frequent health care visits for its management. Individuals without nonemergency medical transportation often miss appointments and do not receive optimal care. This study aims to evaluate the association between Medicaid-provided nonemergency medical transportation and diabetes care visits. A retrospective analysis was conducted of demographic and claims data obtained from the Oklahoma Medicaid program. Participants consisted of Medicaid enrollees with diabetes who made at least 1 visit for diabetes care in a year. The sample was predominantly female and white, with an average age of 46.38 years. Two zero-truncated Poisson regression models were estimated to assess the independent effect of transportation use on number of diabetes care visits. Use of nonemergency medical transportation is a significant predictor of diabetes care visits. Zero-truncated Poisson regression coefficients showed a positive association between the use of transportation and number of visits (0.6563, P < .001). Age, gender, race/ethnicity, area of residence, and presence of additional chronic conditions had independent associations with number of visits. Older enrollees were likely to make more visits than younger enrollees with diabetes (0.02382); controlling for all other factors in the model, rural residents made more visits than urban; women made fewer visits than men (-0.09312; P < .001); and minorities made fewer visits than whites, with pronounced differences for Hispanics and Asians compared to whites. Findings underscore the importance of ensuring transportation to Medicaid populations with diabetes, particularly in the rural areas where the prevalence of diabetes and complications are higher and the availability of medical resources lower than in the urban areas.


Reproductive improvement and senescence in a long-lived bird.
Heterogeneity within a population is a pervasive challenge for studies of individual life-histories. Population-level patterns in age-specific reproductive success can be broken down into relative contributions from selective disappearance, selective appearance of individuals into the study population, and average change in performance for survivors (average ontogenetic development). In this article, we provide an exact decomposition. We apply our formula to data on the reproductive performance of a well characterized population of common terns (Sterna hirundo). We show that improvements with age over most of adult life and senescence at old ages are primarily due to a genuine change in the mean among surviving individuals rather than selective disappearance or selective appearance of individuals. Average ontogenetic development accounts for approximately 87% of the overall age-specific population change.


Cathepsin D activity and selectivity in the acidic conditions of a tumor microenvironment: Utilization in the development of a novel Cathepsin D substrate for simultaneous cancer diagnosis and therapy.
Pro-Cathepsin D (pCD) is an aspartyl endopeptidase which is over expressed in many cancers. This over expression generally led to its secretion into the extracellular culture medium of cancer cells. Moreover, pCD can auto activate and cleave its substrates at an acidic pH compatible with that found in tumor microenvironments (TME). Thus, exploiting these two pathological characteristics of TME offers the opportunity to develop new protease-activated vector on the basis of their specific substrate structures. The aim of this study was to validate new pCD substrates in the extracellular pH conditions of TME. As a first step, we investigated the effect of pH on the catalytic activity and selectivity of mature Cathepsin D (CD). It was found that the increase in the pH of the media led to a decrease in the reaction rate. However, the specificity of mature CD was not affected by a variation in pH. In the second step, the effect of the substrate structure was studied. We demonstrated that the substrate structure had a significant effect on the catalytic activity of CD. In fact, some modifications in peptide structure induced a change in the catalytic behavior that involved a substrate activation phenomenon. We suggest that this activation may be related to the amphiphilic nature of the modified peptide that may induce an interfacial activation mechanism. Finally, pCD, which is the major form found in the extracellular culture medium of cancer cells, was used. We demonstrated that the proform of CD cleave the modified peptide 5 at pH 6.5 with the same cleavage selectivity obtained with the mature form of the protease. These data provide a better understanding of CD behavior in tumor microenvironment conditions and this knowledge can be used to develop more specific tools for diagnosis and drug delivery.


Scanning electron microscopy and evans blue staining for assessing endothelial trauma and reconstitution after venous thrombectomy.
Thrombectomy and a temporary arteriovenous fistula as treatment for deep venous thrombosis has been shown to cause a severe endothelial trauma in an experimental dog study. In vivo staining with Evans blue has been compared to scanning electron microscopy findings to assess the extent of the trauma and the time necessary for endothelial reconstitution. A good correlation between these techniques was found. After the initial almost complete denudation of the veins, new endothelium grew out from small branch openings and valve pockets within four weeks except in the proximity of the arteriovenous fistula where endothelialization was still incomplete after four months.


Conundrums in merging public policy into private dentistry: experiences from Australia's recent past.
Oral disease continues to be a major problem in Australia impacting quality of life, the economy and broader health system. Although the understanding of caries and periodontal disease has improved along with increased government support, oral diseases continue to be the most prevalent among all health conditions. This is despite unprecedented levels of funding in the Chronic Disease Dental Scheme and the Teen Dental Plan. Access to primary care dentistry in the private sector, where the majority of dental services are provided, remains a critical issue. Under the current system of dentistry, it cannot be assumed that the practice of dentistry represents a prioritised approach to combat disease patterns based on scientific evidence in primary health and prevention. Drawing on data in relation to these two programs, the present study highlights issues impacting dental service provision. This includes issues such as access and affordability to dental care, sustainability of policy and its unintended consequences, private practice pressures and the impact of remuneration on treatment. This paper argues that without structural reform there will continue to be barriers in implementing policies capable of improving oral health.


Fluctuating ambient temperature for weaned pigs: effects on performance and immunological and endocrinological functions.
Following weaning at 3 wk of age, crossbred barrows and gilts were housed in temperature-controlled rooms for a 5-d adjustment period at 35 degrees C, then assigned to receive constant ambient temperature (CT) or fluctuating ambient temperature (FT) treatment for the nursery phase of the experiment. Pigs in FT received 12 h at 35 degrees C and 12 h at 15 degrees C daily for the initial 2 wk of the experiment, then 12 h at 29 degrees C and 12 h at 9 degrees C daily for the final 2 wk. Pigs in CT received 35 degrees C and 29 degrees C continuously for the first and final 2 wk, respectively. Weekly growth performance, feed intake and feed-conversion efficiency were not affected by treatment. Plasma glucose, serum cortisol, monocyte phagocytic function and antibody response to a commercial E. coli bacterin were similar in pigs exposed to CT and FT treatments. Concentrations of insulin in serum were similar between treatments at 0, 1 and 3 wk but were increased for pigs in FT at 2 (P less than .05) and 4 wk (P less than .01). Numbers of lymphocytes, band neutrophils and monocytes in pigs were not influenced by ambient temperature treatment. However, numbers of mature neutrophils for pigs in FT were increased (P less than .05) at 1 and 3 wk of treatment. Eosinophils were also elevated in FT pigs at 4 wk of treatment. Pairs of littermate pigs that had been given CT and FT treatments were selected randomly to continue on the finishing phase of the experiment.(ABSTRACT TRUNCATED AT 250 WORDS)


Diabetes Mellitus Predicts Weight Gain After Surgery in Patients With Acromegaly.
Metabolic complications are common in patients with acromegaly. However, this occasionally does not improve post-surgery and may be related to postoperative weight gain. We aimed to investigate the postoperative weight change and factors associated with postoperative weight gain in patients with acromegaly. Overall, 113 consecutive patients with body weight records pre- and 3-6 months post-surgery between October 2009 and March 2021 were enrolled. Patients were divided into three groups: weight loss (weight decrease ≥3%), stable, and weight gain (weight increase ≥3%). Hormone status, metabolic comorbidities, and anthropometric parameters were compared between the groups. Among 113 patients, 29 (25.7%) and 26 (23.0%) patients lost and gained weight, respectively, post-surgery. There were no significant differences in baseline characteristics, including age at diagnosis, sex, body mass index, and growth hormone levels among the three groups. The prevalence of diabetes mellitus at diagnosis was significantly higher in the weight gain group than in the other groups. Patients with diabetes (n=22) had a 5.2-fold higher risk of postoperative weight gain than those with normal glucose tolerance (n=37) (P=0.006). In the diabetes mellitus group, the percentage lean mass decreased (-4.5 [-6.6-2.0]%, P=0.002), and the percentage fat mass significantly increased post-surgery (18.0 [4.6-36.6]%, P=0.003), whereas the normal glucose tolerance group did not show body composition changes post-surgery. In patients with acromegaly, 23% experienced ≥3% weight gain post-surgery. Diabetes mellitus at diagnosis is a significant predictor of weight and fat gain post-surgery.


Type 1 diabetes mellitus and isolated adrenocorticotropin deficiency manifested by parkinsonism: a case report and literature review.
A 67-year-old woman developed isolated adrenocorticotropin deficiency (IAD), which manifested as lethargy, a 20-kg body weight loss, hypoglycemia, and parkinsonism, and began corticosteroid replacement. Her symptoms resolved rapidly, and her weight returned to normal within six months. However, she then developed slowly progressive type 1 diabetes mellitus (T1D) with co-existing Hashimoto thyroiditis, and commenced insulin therapy. To our knowledge, this is the first reported case of parkinsonism associated with IAD. In addition, because diabetes mellitus, including T1D, could be latent in patients with untreated IAD, careful assessment of glucose metabolism is needed after commencing corticosteroid replacement until weight regain is achieved.


Effect of different doses of intrathecal dexmedetomidine on hemodynamic parameters and block characteristics after ropivacaine spinal anesthesia in lower-limb orthopedic surgery: a randomized clinical trial.
The study aims to compare the efficacy of different doses of intrathecal dexmedetomidine on hemodynamic parameters and block characteristics after ropivacaine spinal anesthesia in lower-limb orthopedic surgery. In a double-blind trial, 90 patients undergoing spinal anesthesia for lower-limb orthopedic surgery were included and then randomly assigned to three groups; dexmedetomidine 5 μg/kg, dexmedetomidine 10 μg/kg and placebo. Blood pressure, heart rate, and oxygen saturation were recorded in the three groups at the first 15 minutes and then every 15 to 180 minutes at recovery by a resident anesthesiologist, as well as sensory-motor block onset. The visual analog scale scores for the assessment of pain were recorded at recovery, and 2, 4, 6, and 12 hours postoperatively and the data were analyzed by Stata software. The onset and time to achieve sensory block to ≥ T8 were faster in the 10 μg/kg dexmedetomidine group than the other groups (P = 0.001). The Bromage score was higher in the 10 μg/kg dexmedetomidine group (P = 0.0001) with lower pain score as compared with the 5 μg/kg dexmedetomidine and placebo groups (P = 0.0001). Therefore, an increase in dexmedetomidine hastens the onset of sensory-motor block but not causes side effects. This study was approved by the Ethical Committee of Arak University of Medical Sciences in 2017 (Ethical Code: IR.ARAKMU.REC.1396.37), and the trail was registered and approved by the Iranian Registry of Clinical Trials in 2017 (IRCT No. IRCT2017070614056N12).


Investigation of structure and function of a catalytically efficient variant of the human flavin-containing monooxygenase form 3.
To characterize the contribution of amino acid 360 to the functional activity of the human flavin-containing monooxygenase form 3 (FMO3) and form 1 (FMO1) in the oxygenation of drugs and chemicals, we expressed four FMO3 variants (i.e., Ala360-FMO3, His360-FMO3, Gln360-FMO3, and Pro360-FMO3) and one FMO1 variant (i.e., Pro360-FMO1) and compared them to wild-type enzymes (Leu360-FMO3 and His360-FMO1, respectively). The amino acid substitutions were introduced into wild-type FMO3 or FMO1 cDNA by site-directed mutagenesis. The thermal stability of variants of Leu360 FMO3 was also studied, and the thermal stability was significantly different from that of wild-type FMO3. The influence of different substrates to modulate the catalytic activity of FMO3 variants was also examined. Selective functional substrate activity was determined with mercaptoimidazole, chlorpromazine, and 10-[(N,N-dimethylaminopentyl)-2-(trifluoromethyl)]phenothiazine. Compared with wild-type FMO3, the Ala360-FMO3 and His360-FMO3 variants were less catalytically efficient for mercaptoimidazole S-oxygenation. N-Oxygenation of chlorpromazine was significantly less catalytically efficient for His360-FMO3 compared with wild-type FMO3. Human Pro360-FMO1 was significantly more catalytically efficient at S-oxygenating mercaptoimidazole and chlorpromazine compared with wild-type FMO1. The data support the mechanism that the Pro360 loci affect thermal stability of FMO3. Because different amino acids at position 360 affect substrate oxygenation in a unique fashion compared with that of FMO3 stimulation, we conclude that the mechanism of stimulation of FMO3 is distinct from that of enzyme catalysis. A molecular model of human FMO3 was also constructed to help explain the results. The increase in catalytic efficiency observed for Pro360 in human FMO3 was also observed when the His of FMO1 was replaced by Pro at loci 360.


Robust In Vitro Pharmacology of Tmod, a Synthetic Dual-Signal Integrator for Cancer Cell Therapy.
Progress toward improved solid-tumor treatment has long been hindered by the lack of truly tumor-specific targets. We have developed an approach to T cell therapy based on a dual-receptor system called Tmod™ that addresses this problem. The Tmod system exploits one of the few common genetic differences between tumor and normal cells: loss of heterozygosity (LOH). It utilizes the basic mechanistic logic that evolved in early vertebrates to mediate self vs. non-self discrimination, where an activation stimulus is blocked by self-ligands. Tmod constructs employ a chimeric antigen receptor (CAR) or T cell receptor (TCR) as activator component and a modified LIR-1 inhibitory receptor (blocker) to achieve high selectivity based on expression of the blocker antigen (Ag). Here we explore the in vitro pharmacology of a blocker directed at the HLA-A*02 Ag paired with either a mesothelin CAR or an HLA-A*11-restricted KRAS peptide TCR. While more sensitive to receptor expression changes on effector cells, we show that Tmod response is well-buffered against variations in Ag levels on target cells. In addition, the data reveal at least two distinguishable pharmacologic mechanisms of Tmod blocker function: (1) reducing activator sensitivity and (2) decreasing activation magnitude.


Carbohydrate Conjugates in Vaccine Developments.
Vaccines are powerful tools that can activate the immune system for protection against various diseases. As carbohydrates can play important roles in immune recognition, they have been widely applied in vaccine development. Carbohydrate antigens have been investigated in vaccines against various pathogenic microbes and cancer. Polysaccharides such as dextran and β-glucan can serve as smart vaccine carriers for efficient antigen delivery to immune cells. Some glycolipids, such as galactosylceramide and monophosphoryl lipid A, are strong immune stimulators, which have been studied as vaccine adjuvants. In this review, we focus on the current advances in applying carbohydrates as vaccine delivery carriers and adjuvants. We will discuss the examples that involve chemical modifications of the carbohydrates for effective antigen delivery, as well as covalent antigen-carbohydrate conjugates for enhanced immune responses.


Connectomics of synaptic microcircuits: lessons from the outer retina.
Photoreceptors form a sophisticated synaptic complex with bipolar and horizontal cells, transmitting the signals generated by the phototransduction cascade to downstream retinal circuitry. The cone photoreceptor synapse shows several characteristic anatomical connectivity motifs that shape signal transfer: typically, ON-cone bipolar cells receive photoreceptor input through invaginating synapses; OFF-cone bipolar cells form basal synapses with photoreceptors. Both ON- and OFF-cone bipolar cells are believed to sample from all cone photoreceptors within their dendritic span. Electron microscopy and immunolabelling studies have established the robustness of these motifs, but have been limited by trade-offs in sample size and spatial resolution, respectively, constraining precise quantitative investigation to a few individual cells. 3D-serial electron microscopy overcomes these limitations and has permitted complete sets of neurons to be reconstructed over a comparatively large section of retinal tissue. Although the published mouse dataset lacks labels for synaptic structures, the characteristic anatomical motifs at the photoreceptor synapse can be exploited to identify putative synaptic contacts, which has enabled the development of a quantitative description of outer retinal connectivity. This revealed unexpected exceptions to classical motifs, including substantial interaction between rod and cone pathways at the photoreceptor synapse, sparse photoreceptor sampling and atypical contacts. Here, we summarize what was learned from this study in a more general context: we consider both the implications and limitations of the study and identify promising avenues for future research.


A prospective multicentre controlled study of Gaoweikang (Chinese multiherb extract-based tincture) used in high-risk HPV infections.
The aim of the study was to investigate the safety and antiviral efficacy of a Chinese multiherb extract-based tincture (GWK) on a population of patients with high-risk human papilloma (hrHPV) infections and hrHPV-caused cervical low-grade squamous intraepithelial lesions (LSILs). Patients with persistent hrHPV infection were enrolled in Group A, including A1 subjects, who received the intervention, and A2 subjects, who received the control. Patients with hrHPV infection causing cervical LSIL were enrolled in Group B, which included B1 subjects, who received the intervention, and B2 subjects, who served as the control. For Groups A1 and B1, hrHPV was tested at 3 months (M3) and 6 months (M6) after the intervention. The side effects were also analyzed. At baseline (D0), a total of 99 patients were enrolled in Group A, with 50 subjects in Group A1 and 49 subjects in Group A2. A total of 91 patients were enrolled in Group B, with 45 subjects in Group B1 and 46 subjects in Group B2. There was no significant difference in the characteristics, including average age, age stratification, and HPV genotype. At M6, both Group A1 and Group B1 had a higher hrHPV clearance rate than the control group (A1/A2: 80.0% vs. 20.4%; B1/B2: 64.4% vs. 15.2%, p<0.001). At M6, the effective rates of Group A1 and Group B1 were 84% (42/50) and 68.9% (31/45), respectively. The side effect rates of Groups A1 and B1 were 11.5% (6/52) and 11.1% (5/45), respectively. Most adverse reactions involved local discomfort, including vulvar erythema, vulvar itch, increased vaginal discharge, cervical bleeding, and mild pain in the lower abdomen. Univariate logistic regression analysis showed that the intervention had an OR of 12 (95% CI 4.431-32.50) for clearing persistent HPV infection (p<0.001). For cervical LSIL, the intervention had an OR of 10.1 for clearing persistent HPV infection (95% CI 3.68-27.7) (p<0.001). The results of this study suggest that the Chinese multiherb extract-based tincture GWK is safe and well tolerated. Furthermore, this preliminary study showed that this Chinese multiherb extract-based tincture is helpful for promoting HPV clearance in cases of persistent HPV and HPV-induced LSIL.


Cardiac regeneration: All work and no repair?
Structural changes in the developing heart may influence the limited regenerative capacity of the adult heart. We examine how the workload exerted on the adult mammalian heart may limit regenerative capability and discuss recent therapies that demonstrate beneficial effects through unloading the heart.


Comparison of the biotransformation of the 14C-labelled insecticide carbaryl by non-transformed and human CYP1A1-, CYP1A2-, and CYP3A4-transgenic cell cultures of Nicotiana tabacum.
Transgenic tobacco-cell-suspension cultures expressing separately the human cytochrome P450 monooxygenases CYP1A1, CYP1A2, and CYP3A4 were utilized to study the biotransformation of the 14C-labelled insecticide carbaryl (=naphthalen-1-yl methylcarbamate). The resulting data were compared to similar data from the corresponding non-transformed (NT) tobacco-cell culture and commercially available membrane preparations (Bactosomes) of genetically modified bacteria separately containing the same human P450s. A rapid conversion rate of carbaryl was observed with the CYP1A1 and CYP1A2 cells, where only 49.7 and 0.2% of applied carbaryl (1 mg/l), respectively, remained after 24 h, as compared to 77.7% in the non-transformed culture. Unexpectedly, the corresponding results obtained from the CYP3A4 cultures were not definite. With 25 mg/l of carbaryl and 96 h of incubation, it was proven that the insecticide is also substrate of CYP3A4. This finding was supported by GC/EI-MS analysis of the primary metabolite pattern produced by the isozyme. This consisted of naphthalene-1-ol, N-(hydroxymethyl)carbaryl, 4-hydroxycarbaryl, and 5-hydroxycarbaryl, whereas the main product in non-transformed cells was N-(hydroxymethyl)carbaryl. Data obtained from the CYP1A1, CYP1A2, or CYP3A4 Bactosomes agreed with those of the P450-transgenic tobacco cells. Problems with GC/EI-MS analysis of carbaryl and its metabolites are discussed.


Extended release of a large amount of highly water-soluble diltiazem hydrochloride by utilizing counter polymer in polyethylene oxides (PEO)/polyethylene glycol (PEG) matrix tablets.
The purpose of this study was to evaluate the feasibility of using a counter polymer in polyethylene oxide (PEO)/polyethylene glycol (PEG) polymeric matrices for the sustained release of a large amount of highly water-soluble drug. PEO/PEG matrix tablets (CR-A) containing four drugs with different water solubilities were prepared to investigate the effect of drug solubility on the drug-release and diffusion properties of PEO/PEG matrices. Cross-linked carboxyvinyl polymer (CVP)/PEO/PEG matrix tablets (CR-B) containing a water-soluble drug, diltiazem hydrochloride (DTZ), were also prepared, and their in vitro characteristics were compared with those of CR-A. Their in vitro drug release properties were evaluated using a dissolution test, and the polymeric erosion and drug diffusion properties of the matrices were also calculated. Drugs with higher solubility in water were released faster for the CR-A. The drug-release rate also increased with the amount of drug loaded. CR-A containing 50% DTZ (by weight) extended drug release by only 6h. This confirms the difficulty experienced when trying to formulate PEO/PEG matrices for the sustained release of a large amount of highly water-soluble drugs due to large drug diffusion. In an attempt to control this issue, a polymer bearing a charge opposite that of the drug was used to effectively decrease the diffusion of DTZ, resulting in sustained release for 24h or longer. These results suggested that including counter polymer in the PEO/PEG matrix tablet is a useful tool for achieving the sustained release of a large amount of highly water-soluble drug.


RhoD participates in the regulation of cell-cycle progression and centrosome duplication.
We have previously identified a Rho protein, RhoD, which localizes to the plasma membrane and the early endocytic compartment. Here, we show that a GTPase-deficient mutant of RhoD, RhoDG26V, causes hyperplasia and perturbed differentiation of the epidermis, when targeted to the skin of transgenic mice. In vitro, gain-of-function and loss-of-function approaches revealed that RhoD is involved in the regulation of G1/S-phase progression and causes overduplication of centrosomes. Centriole overduplication assays in aphidicolin-arrested p53-deficient U2OS cells, in which the cell and the centrosome cycles are uncoupled, revealed that the effects of RhoD and its mutants on centrosome duplication and cell cycle are independent. Enhancement of G1/S-phase progression was mediated via Diaph1, a novel effector of RhoD, which we have identified using a two-hybrid screen. These results indicate that RhoD participates in the regulation of cell-cycle progression and centrosome duplication.


XRCC3 gene polymorphism is associated with survival in Japanese lung cancer patients.
We focused on OGG1 Ser326Cys, MUTYH Gln324His, APEX1 Asp148Glu, XRCC1 Arg399Gln, and XRCC3 Thr241Met and examined the relationship between the different genotypes and survival of Japanese lung cancer patients. A total of 99 Japanese lung cancer patients were recruited into our study. Clinical data were collected, and genotypes of the target genes were identified by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Survival analysis to verify the impact of these gene polymorphisms on the clinical outcome of lung cancer showed that lung squamous cell carcinoma patients with the Thr/Met genotype at XRCC3 had a significantly shorter survival time than those with the Thr/Thr genotype (13 months versus 48 months; log-rank test, p < 0.0001). Cox regression analysis showed that the carriers of XRCC3 genotypes were at a significantly higher risk [adjusted hazard ratio (HR) = 9.35, 95% confidence interval (CI) = 2.52-34.68, p = 0.001; adjusted HR = 9.05, 95% CI = 1.89-44.39, p = 0.006]. Our results suggest that XRCC3 Thr241Met may act as a favorable prognostic indicator for lung squamous cell carcinoma patients.