Combination of lidocaine suppository and periprostatic nerve block during transrectal prostate biopsy: a prospective randomized trial.

OBJECTIVE: To assess the most effective local analgesia during transrectal ultrasound-guided prostate biopsy. METHODS: A total of 123 consecutive patients undergoing transrectal ultrasound-guided prostate biopsy for elevated prostate-specific antigen levels and/or a suspicious digital rectal examination were randomized to three groups. Patients received a 60-mg lidocaine suppository (group 1, n = 41), a periprostatic nerve block (10-mL injection of lidocaine hydrochloride; group 2, n = 41) or a combination of both (group 3, n = 41) before a 10-core transrectal ultrasound-guided biopsy. A total of 80.5% (n = 99) of the patients underwent their first biopsy, 27.1% (n = 22) their second and 2.4% (n = 2) the third. Pain was evaluated on a 10-point visual analog scale for each step of the procedure. RESULTS: Prostate-specific antigen values ranged from 0.39 to 90.1 (mean [SD] 8.76 ng/mL [11.08 ng/mL]). Comparison of the median visual analog scale scores between groups 1 and 2 showed a significant difference (P = 0.004). The differences in the outcomes between groups 1 and 3 (P = 0.001), and groups 2 and 3 (P = 0.001) were also significant. Patients of group 3 had the best output corresponding to the pain sensations and therefore the lowest visual analog scale scores. CONCLUSION: The combination of lidocaine suppository and periprostatic lidocaine infiltration is more effective for pain control than either lidocaine suppository or periprostatic lidocaine infiltration alone in patients undergoing transrectal ultrasound-guided prostate biopsy.

DNA extraction from clotted blood in genotyping quality.

DNA extraction from frozen blood clots is challenging. Here, the authors applied QIAGEN Clotspin Baskets and the Gentra Puregene Blood Kit for DNA extraction to cellular fraction of 5.5 ml whole blood without anticoagulating additives. The amount and quality of extracted DNA were assessed via spectrophotometer and gel electrophoresis. Results from array-based genotyping were analyzed. All steps were compared with DNA isolated from anticoagulated blood samples from a separate study. The quality and concentration of DNA extracted from clotted blood were comparable to those of DNA extracted from anticoagulated blood. DNA yield was on average 27 µg per ml clotted blood, with an average purity of 1.87 (A260/A280). Genotyping quality was similar for both DNA sources (call rate: 99.56% from clotted vs 99.49% from anticoagulated blood).

Structural basis for TBP displacement from TATA box DNA by the Swi2/Snf2 ATPase Mot1.

The Swi2/Snf2 family transcription regulator Modifier of Transcription 1 (Mot1) uses adenosine triphosphate (ATP) to dissociate and reallocate the TATA box-binding protein (TBP) from and between promoters. To reveal how Mot1 removes TBP from TATA box DNA, we determined cryogenic electron microscopy structures that capture different states of the remodeling reaction. The resulting molecular video reveals how Mot1 dissociates TBP in a process that, intriguingly, does not require DNA groove tracking. Instead, the motor grips DNA in the presence of ATP and swings back after ATP hydrolysis, moving TBP to a thermodynamically less stable position on DNA. Dislodged TBP is trapped by a chaperone element that blocks TBP's DNA binding site. Our results show how Swi2/Snf2 proteins can remodel protein-DNA complexes through DNA bending without processive DNA tracking and reveal mechanistic similarities to RNA gripping DEAD box helicases and RIG-I-like immune sensors.

microRNA165 and 166 modulate response of the Arabidopsis root apical meristem to salt stress.

In plants, developmental plasticity allows for the modulation of organ growth in response to environmental cues. Being in contact with soil, roots are the first organ that responds to various types of soil abiotic stress such as high salt concentration. In the root, developmental plasticity relies on changes in the activity of the apical meristem, the region at the tip of the root where a set of self-renewing undifferentiated stem cells sustain growth. Here, we show that salt stress promotes differentiation of root meristem cells via reducing the dosage of the microRNAs miR165 and 166. By means of genetic, molecular and computational analysis, we show that the levels of miR165 and 166 respond to high salt concentration, and that miR165 and 166-dependent PHABULOSA (PHB) modulation is central to the response of root growth to this stress. Specifically, we show that salt-dependent reduction of miR165 and 166 causes a rapid increase in PHB expression and, hence, production of the root meristem pro-differentiation hormone cytokinin. Our data provide direct evidence for how the miRNA-dependent modulation of transcription factor dosage mediates plastic development in plants.

A PHABULOSA-Controlled Genetic Pathway Regulates Ground Tissue Patterning in the Arabidopsis Root.

In both animals and plants, development involves anatomical modifications. In the root of Arabidopsis thaliana, maturation of the ground tissue (GT)-a tissue comprising all cells between epidermal and vascular ones-is a paradigmatic example of these modifications, as it generates an additional tissue layer, the middle cortex (MC).1-4 In early post-embryonic phases, the Arabidopsis root GT is composed of one layer of endodermis and one of cortex. A second cortex layer, the MC, is generated by asymmetric cell divisions in about 80% of Arabidopsis primary roots, in a time window spanning from 7 to 14 days post-germination (dpg). The cell cycle regulator CYCLIN D6;1 (CYCD6;1) plays a central role in this process, as its accumulation in the endodermis triggers the formation of MC.5 The phytohormone gibberellin (GA) is a key regulator of the timing of MC formation, as alterations in its signaling and homeostasis result in precocious endodermal asymmetric cell divisions.3,6,7 However, little is known on how GAs are regulated during GT maturation. Here, we show that the HOMEODOMAIN LEUCINE ZIPPER III (HD-ZIPIII) transcription factor PHABULOSA (PHB) is a master regulator of MC formation, controlling the accumulation of CYCD6;1 in the endodermis in a cell non-autonomous manner. We show that PHB activates the GA catabolic gene GIBBERELLIN 2 OXIDASE 2 (GA2ox2) in the vascular tissue, thus regulating the stability of the DELLA protein GIBBERELLIN INSENSITIVE (GAI)-a GA signaling repressor-in the root and, hence, CYCD6;1 expression in the endodermis.

Are "normal" multidetector computed tomographic scans sufficient to allow collar removal in the trauma patient?

BACKGROUND: Controversy continues as to the most safe and reliable method for clearing the cervical spine (C-spine) in a trauma patient who is rendered unable to participate in a clinical examination. Although magnetic resonance imaging (MRI) is the most sensitive test to detect soft-tissue injuries, it is impractical for routine use in every patient largely because of its cost and time of acquiescence. Recent studies have advocated the sole use of multidetector computed tomographic (MDCT) scans of the C-spine to decide if cervical collar immobilization can be discontinued. The current investigation retrospectively reviewed a series of MDCT scans obtained after an acute traumatic event that were used to direct treatment in the emergency department (ED) or intensive care unit. METHODS: Seven-hundred and eight trauma patients consecutively admitted to the ED between June 2001 and July 2006 underwent a computed tomographic scan of their C-spine as part of an institutional protocol. We identified 91 patients with MDCT scans that were officially recorded as adequate and negative by an attending ED radiologist who had also undergone an MRI during the same trauma admission period. Retrospectively, two fellowship-trained spine surgeons independently reviewed these MDCT studies to address the following questions: (1) Is the study adequate? (2) Is it suggestive of an acute injury? (3) Is there sufficient information to safely recommend collar removal? Institutional Review Board approval was obtained before the images were reviewed. Neither clinical examination findings nor MRI readings were made available to the surgeon evaluators. RESULTS: Both spine surgeons agreed that 76 of the 91 studies (84%) were adequate to evaluate for possible C-spine injuries. Seven of 91 MDCT scans (8%) were deemed inadequate by both surgeons (95% confidence interval, 2.3-13.1). Reasons for inadequacy included motion artifact, insufficient visualization of the cervical-thoracic or occipital-cervical junctions, incomplete reconstructive views, or poor quality. Three of the adequate MDCT scans had fractures that were identified by both of the spine surgeons; 4 additional fractures and 15 findings suspicious for instability were identified by at least one of the surgeons. Ultimately, 22 of 91 MDCT scans read as adequate and normal by attending radiologists were deemed suspicious for abnormality by the spine surgeons. Of these 22 cases, the official MRI reading was positive for a trauma-related abnormality in 17 cases. CONCLUSIONS: C-spine clearance of patients without the ability participate in a clinical examination remains difficult. A multidisciplinary, algorithmic approach generally yields the most consistent results. However, our data highlight that reliance on a single imaging modality may lead to missed diagnosis of C-spine injuries. These data suggest that early involvement of the spine service for radiographic clearance may help identify occult injuries or suspicious findings necessitating further evaluation.

Cytokinin-Dependent Control of GH3 Group II Family Genes in the Arabidopsis Root.

Abstract: The Arabidopsis root is a dynamic system where the interaction between different plant hormones controls root meristem activity and, thus, organ growth. In the root, a characteristic graded distribution of the hormone auxin provides positional information, coordinating the proliferating and differentiating cell status. The hormone cytokinin shapes this gradient by positioning an auxin minimum in the last meristematic cells. This auxin minimum triggers a cell developmental switch necessary to start the differentiation program, thus, regulating the root meristem size. To position the auxin minimum, cytokinin promotes the expression of the IAA-amido synthase group II gene GH3.17, which conjugates auxin with amino acids, in the most external layer of the root, the lateral root cap tissue. Since additional GH3 genes are expressed in the root, we questioned whether cytokinin to position the auxin minimum also operates via different GH3 genes. Here, we show that cytokinin regulates meristem size by activating the expression of GH3.5 and GH3.6 genes, in addition to GH3.17. Thus, cytokinin activity provides a robust control of auxin activity in the entire organ necessary to regulate root growth.

Alkenylation of unactivated alkyl bromides through visible light photocatalysis.

Two visible-light driven alkenylation reactions of unactivated alkyl bromides, which were enabled by the use of Ir(dF(CF3)ppy)2(dtbbpy)PF6 as the photocatalyst and (TMS)3SiH as the atom transfer reagent to activate the alkyl bromides, were described for the first time. These protocols can be used to produce a variety of alkenes from easily available feedstock with good reaction efficiency and high chemoselectivity under mild reaction conditions. To further demonstrate the applicability of the present strategy, the alkenylation of bioactive molecules and glycosyl bromides, as well as the alkynylation of unactivated alkyl bromides, was proven to be feasible.

Interplay between phosphorylation and SUMOylation events determines CESTA protein fate in brassinosteroid signalling.

Brassinosteroids (BRs) are steroid hormones that are essential for plant growth. Responses to these hormones are mediated by transcription factors of the bri1-EMS suppressor 1/brassinazole resistant 1 subfamily, and BRs activate these factors by impairing their inhibitory phosphorylation by GSK3/shaggy-like kinases. Here we show that BRs induce nuclear compartmentalization of CESTA (CES), a basic helix-loop-helix transcription factor that regulates BR responses, and reveal that this process is regulated by CES SUMOylation. We demonstrate that CES contains an extended SUMOylation motif, and that SUMOylation of this motif is antagonized by phosphorylation to control CES subnuclear localization. Moreover, we provide evidence that phosphorylation regulates CES transcriptional activity and protein turnover by the proteasome. A coordinated modification model is proposed in which, in a BR-deficient situation, CES is phosphorylated to activate target gene transcription and enable further posttranslational modification that controls CES protein stability and nuclear dynamics.

Brigham fracture intervention team initiatives for hospital patients with hip fractures: a paradigm shift.

We designed, implemented, and revised the Brigham Fracture Intervention Team (B-FIT) initiatives to improve in-hospital care of fracture (Fx) patients. Effectiveness was evaluated for 181 medical records of 4 cohorts in four successive years of consecutive patients who were admitted with a fragility hip Fx. The Discharge Initiative (DI) (computer-based) includes 1200 mg calcium and 1000 IU vitamin D(3) daily. The Admission Initiative (AI) was introduced one year later with reminders for serum 25OHD measurement, initiation of daily calcium (1200 mg) and vitamin D (800 IU), and an order for Endocrinology consultation, with an amendment for a computer-assisted reminder and a dose of D(2) (50 000 IU). Initially, the computer-based DI was more effective (67%) than the surgeon-driven AI (33%, P < .001). After introduction of a computer-assisted reminder, AI effectiveness increased to 68%. The marked prevalence of vitamin D insufficiency reaffirms the importance of incorporating vitamin D recommendations in Fx care pathways.

Mortality in elderly patients after cervical spine fractures.

BACKGROUND: Despite an increased risk of cervical spine fractures in older patients, little is known about the mortality associated with these fractures and there is no consensus on the optimal treatment. The purposes of this study were to determine the three-month and one-year mortality associated with cervical spine fractures in patients sixty-five years of age or older and to evaluate potential factors that may influence mortality. METHODS: We performed a retrospective review of all cervical spine fractures in patients sixty-five years of age or older from 1991 to 2006 at two institutions. Information regarding age, sex, race, treatment type, neurological involvement, injury mechanism, comorbidity, and mortality were collected. Overall risk of mortality and mortality stratified by the above factors were calculated at three months and one year. Cox proportional-hazard regression was performed to identify independent correlates of mortality. RESULTS: Six hundred and forty patients were included in our analysis. The mean age was eighty years (range, sixty-five to 101 years). Two hundred and ninety-four patients (46%) were male, and 116 (18%) were nonwhite. The risk of mortality was 19% at three months and 28% at one year. The effect of treatment on mortality varied with age at three months (p for interaction = 0.03) but not at one year (p for interaction = 0.08), with operative treatment being associated with less mortality for those between the ages of sixty-five and seventy-four years. A higher Charlson comorbidity score, male sex, and neurological involvement were all associated with increased risk of mortality. CONCLUSIONS: Operative treatment of cervical spine fractures is associated with a lower mortality rate at three months but not at one year postoperatively for patients between sixty-five and seventy-four years old at the time of fracture.