CRISPRi-based circuits to control gene expression in plants.

The construction of synthetic gene circuits in plants has been limited by a lack of orthogonal and modular parts. Here, we implement a CRISPR (clustered regularly interspaced short palindromic repeats) interference (CRISPRi)-based reversible gene circuit platform in plants. We create a toolkit of engineered repressible promoters of different strengths and construct NOT and NOR gates in Arabidopsis thaliana protoplasts. We determine the optimal processing system to express single guide RNAs from RNA Pol II promoters to introduce NOR gate programmability for interfacing with host regulatory sequences. The performance of a NOR gate in stably transformed Arabidopsis plants demonstrates the system's programmability and reversibility in a complex multicellular organism. Furthermore, cross-species activity of CRISPRi-based logic gates is shown in Physcomitrium patens, Triticum aestivum and Brassica napus protoplasts. Layering multiple NOR gates together creates OR, NIMPLY and AND logic functions, highlighting the modularity of our system. Our CRISPRi circuits are orthogonal, compact, reversible, programmable and modular and provide a platform for sophisticated spatiotemporal control of gene expression in plants.

Systematic review and meta-analysis comparing Manta device and Perclose device for closure of large bore arterial access.

Data comparing MANTA device with Perclose device for large bore arterial access closure is limited. We performed meta-analysis to compare safety and efficacy of the two devices in large (⩾14 Fr sheath) arteriotomy closure post-TAVR. Relevant studies were identified via PubMed, Cochrane, and EMBASE databases until June, 2022. Data was analyzed using random effect model to calculate relative odds of VARC-2 defined access-site complications and short-term (in-hospital or 30-day) mortality. A total of 12 studies (2 RCT and 10 observational studies) comprising 2339 patients were included. The odds of major vascular complications (OR 0.99, 95% CI 0.51-1.92; p = 0.98); life threatening and major bleeding (OR 0.77, 95% CI 0.45-1.33; p = 0.35); minor vascular complications (OR 1.37, 95% CI 0.63-2.99; p = 0.43); minor bleeding (OR 0.94, 95% CI 0.57-1.56; p = 0.82); device failure (OR 0.74, 95% CI 0.49-1.11; p = 0.14); hematoma formation (OR 0.76, 95% CI 0.33-1.75; p = 0.52); dissection, stenosis, occlusion, or pseudoaneurysm (OR 1.08, 95% CI 0.71-1.62; p = 0.73) and short-term mortality (OR 1.01, 95% CI 0.55-1.84; p = 0.98) between both devices were similar. MANTA device has a similar efficacy and safety profile compared to Perclose device.

A Single Stage Composite Cleft Septorhinoplasty for Correction of the Mature Unilateral Cleft Nose Deformity - The Gujrat Technique.

OBJECTIVE: To evaluate the results of a single stage composite cleft septorhinoplasty procedure ("The Gujrat Technique") to correct the exaggerated cleft nose deformity after completion of nasal growth in an adult patient cohort. METHODS: Adult patients with a residual unilateral cleft nasal deformity were deemed eligible for the proposed "Gujrat Technique". Over a 10-year period (2007-2017), 96 adult patients underwent this composite cleft septorhinoplasty as a single stage operation. Post-operative nasal symmetry evaluation was undertaken using the validated computer program 'SymNose'. Functional outcome and patient satisfaction were assessed using Nasal Obstruction Symptom Evaluation scale and Rhinoplasty Outcome Evaluation (ROE) questionnaires respectively. Various statistical analysis methods were used to validate the obtained results. RESULTS: Due to poor compliance with follow-up, post-operative assessments were undertaken in only 32 patients. The single group study design using the non-parametric matching pairs Wilcoxon Sign test (p < 0.001) showed overall good to excellent functional and aesthetic outcomes and higher scores of the digital SymNose grading system. There was a significant improvement in ROE scores (from 26.4 ± 2.9 to 85.9 ± 4.7, p < 0.001). There were no major complications or revisions needed in our series. CONCLUSION: The individual components of "The Gujrat Technique" are not novel but their combination in this adult unilateral cleft rhinoplasty cohort has demonstrated a high patient satisfaction with its aesthetic appeal and functional versatility. In the background of limited resources and unpredictable patient follow up, the simplicity, reproducibility and cost effectiveness of this technique make it a practical reconstructive option.

Smoking and Fibrocystic Changes in the Breast: A Case Report of a Lifelong Smoker and Changes in Breast Parenchyma.

Smoking is a well-known risk factor for breast cancer, and the nicotine in cigarette smoke has been associated with fibrotic changes in the breast. Although considered benign, these changes have the potential to make surgical dissection more difficult and may increase the risk of surgical complications. Here we discuss the case of a middle-aged female who was a lifelong smoker with breast cancer and who underwent a simple mastectomy. Intra-operatively, the overall breast parenchyma appeared quite firm, making tissue dissection challenging and the operating surgeon had to endure to remove the tumor and the remaining breast tissue.

Synthetic memory circuits for stable cell reprogramming in plants.

Plant biotechnology predominantly relies on a restricted set of genetic parts with limited capability to customize spatiotemporal and conditional expression patterns. Synthetic gene circuits have the potential to integrate multiple customizable input signals through a processing unit constructed from biological parts to produce a predictable and programmable output. Here we present a suite of functional recombinase-based gene circuits for use in plants. We first established a range of key gene circuit components compatible with plant cell functionality. We then used these to develop a range of operational logic gates using the identify function (activation) and negation function (repression) in Arabidopsis protoplasts and in vivo, demonstrating their utility for programmable manipulation of transcriptional activity in a complex multicellular organism. Specifically, using recombinases and plant control elements, we activated transgenes in YES, OR and AND gates and repressed them in NOT, NOR and NAND gates; we also implemented the A NIMPLY B gate that combines activation and repression. Through use of genetic recombination, these circuits create stable long-term changes in expression and recording of past stimuli. This highly compact programmable gene circuit platform provides new capabilities for engineering sophisticated transcriptional programs and previously unrealized traits into plants.

Exploring the Perceptions Surrounding Hospital Chaplains in Patient Care and Healing.

The authors distributed a questionnaire to various hospital staff to explore perceptions surrounding the role of chaplains in patient care and healing through assessing opinions, beliefs, and knowledge. Results revealed a poor understanding of the role of the hospital chaplain in the UK. The authors therefore advocate education programs to increase awareness of the varied role of the hospital chaplain in fulfilling patient spiritual needs to increase referral practices, leading to improved patient outcomes.

IV3000 semi-occlusive dressing use in simple and complex fingertip injuries: efficacy and affordability.

OBJECTIVE: This study aimed to explore the efficacy of the IV3000 semi-occlusive, transparent adhesive film dressing in the non-surgical management of simple as well as more complex fingertip injuries. METHOD: In this qualitative study, patients with fingertip injuries were prospectively recruited and treated conservatively with the dressing between 2015 and 2017. Inclusion criteria included any fingertip injury with tissue loss and patient consent for non-surgical treatment consistent with the study protocol. Exclusion criteria included injuries needing surgical intervention for tendon injury or exposure, joint dislocations, distal phalangeal fractures requiring fixation, bone exposure, isolated nail bed lacerations and any patients eligible for surgical repair who did not wish to be managed conservatively. RESULTS: A total of 64 patients took part in the study. The patients treated with the dressing were asked to rate functional outcome, of whom 40 (62.5%) patients reported the outcome as 'excellent', 19 (29.7%) as 'satisfactory', five (7.8%) as 'indifferent' and none (0%) as 'unsatisfactory'. A reduced pulp volume at completion of healing was felt by 21 (32.8%) patients, but all patients were 'satisfied' with the aesthetic appearance of their fingertips at final clinical review. Average healing time was 4.5 weeks across the group, with the average time for return to work being just under one week. We estimate a 60% reduction in cost with the conservative versus the surgical management option. CONCLUSION: This study showed that, for participants, the IV3000 dressing was an affordable and effective option for the conservative treatment of simple fingertip injuries and in the management of more complex fingertip injuries.

Design and Analysis of In-Pipe Hydro-Turbine for an Optimized Nearly Zero Energy Building.

Pakistan receives Direct Normal Irradiation (DNI) exceeding 2000 kWh/m²/annum on approximately 83% of its land, which is very suitable for photovoltaic production. This energy can be easily utilized in conjunction with other renewable energy resources to meet the energy demands and reduce the carbon footprint of the country. In this research, a hybrid renewable energy solution based on a nearly Zero Energy Building (nZEB) model is proposed for a university facility. The building in consideration has a continuous flow of water through its water delivery vertical pipelines. A horizontal-axis spherical helical turbine is designed in SolidWorks and is analyzed through a computational fluid dynamics (CFD) analysis in ANSYS Fluent 18.1 based on the K-epsilon turbulent model. Results obtained from ANSYS Fluent have shown that a 24 feet vertical channel with a water flow of 0.2309 m3/s and velocity of 12.66 m/s can run the designed hydroelectric turbine, delivering 168 W of mechanical power at 250 r.p.m. Based on the turbine, a hybrid renewable energy system (HRES) comprising photovoltaic and hydroelectric power is modelled and analyzed in HOMER Pro software. Among different architectures, it was found that architecture with hydroelectric and photovoltaic energy provided the best COE of $0.09418.

Atypical Fibroxanthoma Within a Melanoma: A Case Report.

The finding of a pigmented lesion within another distinct lesion is rare but not unheard of. Here, we describe the presence of an atypical fibroxanthoma within a melanoma in a 72-year-old female referred to the plastics surgery department with a pigmented lesion on her left knee. It was excised in view of clinical suspicion of melanoma. The histopathology report documented a single lesion with two distinct components, namely a melanoma of superficial spreading type with a Breslow thickness of 3.0mm, and a central nodule of atypical fibroxanthoma.

Paediatric Burns From Deployment of a Concealed Aviation Seatbelt Airbag.

The advantages of airbags in reducing the rate of severe injuries and fatalities in motor vehicle crashes are well known but the physical act of airbag deployment can lead to injury to the passenger and the spectrum of airbag trauma resulting from deployment of vehicle airbags has been extensively reported. We present the first reported case of a pediatric burn injury resulting from the accidental deployment of an airbag in an aircraft. A four-year-old female child sustained injuries to the left side of her face and body while she was aboard a stationary airplane and the airbag concealed within the seat belt of her airplane seat unexpectedly and inexplicably deployed just before departure. We are presenting the case to increase awareness of the possibility of this injury in aircraft and to enable minimization of such accidents as well as help establish protocols for dealing with such mishaps if there were to happen.

Subcutaneous fat necrosis requiring plastic surgical intervention in an infant treated with whole-body cooling.

We report an infant who developed subcutaneous fat necrosis of the newborn (SCFN) secondary to cooling treatment for hypoxic ischaemic encephalopathy (HIE). While SCFN is usually self-limiting, this patient went on to develop a large haematoma on his back with overlying skin necrosis necessitating debridement and split thickness skin grafting. Initially, the area affected on his back showed a number of small fluctuant swellings. By day 16 after birth, theses swellings coalesced to form a large 15 cm × 19 cm haematoma with a tense, shiny skin overlying it. On day 17, the large swelling was drained in theatre and a drain was left in situ. Total calcium blood level was raised at 4 mmol/l and he was managed with Pamidronate infusion. Postoperatively, examination of the back showed a 5 cm necrotic area in the centre of the back, and affected area was debrided along with a split skin graft applied to the exposed area.

Genome analysis and avirulence gene cloning using a high-density RADseq linkage map of the flax rust fungus, Melampsora lini.

BACKGROUND: Rust fungi are an important group of plant pathogens that cause devastating losses in agricultural, silvicultural and natural ecosystems. Plants can be protected from rust disease by resistance genes encoding receptors that trigger a highly effective defence response upon recognition of specific pathogen avirulence proteins. Identifying avirulence genes is crucial for understanding how virulence evolves in the field. RESULTS: To facilitate avirulence gene cloning in the flax rust fungus, Melampsora lini, we constructed a high-density genetic linkage map using single nucleotide polymorphisms detected in restriction site-associated DNA sequencing (RADseq) data. The map comprises 13,412 RADseq markers in 27 linkage groups that together span 5860 cM and contain 2756 recombination bins. The marker sequences were used to anchor 68.9 % of the M. lini genome assembly onto the genetic map. The map and anchored assembly were then used to: 1) show that M. lini has a high overall meiotic recombination rate, but recombination distribution is uneven and large coldspots exist; 2) show that substantial genome rearrangements have occurred in spontaneous loss-of-avirulence mutants; and 3) identify the AvrL2 and AvrM14 avirulence genes by map-based cloning. AvrM14 is a dual-specificity avirulence gene that encodes a predicted nudix hydrolase. AvrL2 is located in the region of the M. lini genome with the lowest recombination rate and encodes a small, highly-charged proline-rich protein. CONCLUSIONS: The M. lini high-density linkage map has greatly advanced our understanding of virulence mechanisms in this pathogen by providing novel insights into genome variability and enabling identification of two new avirulence genes.

Pediatric Traumatic Limb Amputation: The Principles of Management and Optimal Residual Limb Lengths.

Pediatric traumatic limb amputations are rare and their acute and long term management can be challenging in this subgroup of patients. The lengthy and costly hospital stays, and resulting physical and psychological implications leads to significant morbidity. We present a summary of treatment principles and the evidence base supporting the management options for this entity. The initial management focuses on resuscitating and stabilization of the patients, administration of appropriate and adequate analgesics, and broad spectrum antibiotics. The patient should ideally be managed by an orthopedic or a plastic surgeon and when an amputation is warranted, the surgical team should aim to conserve as much of the viable physis as possible aimed at allowing bone development in a growing child. A subsequent wound inspection should be performed to assess for signs of ischemia or non-viability of tissue. Depending on the child's age, approximations of the ideal residual limb length can be calculated using our guidelines, allowing an ideal stump length at skeletal maturity for a well-fitting and appropriate prosthesis. Myodesis and myoplasties can be performed according to the nature of the amputation. Removable rigid dressings are safe and cost effective offering better protection of the stump. Complications such as necrosis and exostosis, on subsequent examination, warrant further revisions. Other complications such as neuromas can be prevented by proximal division of the nerves. Successful rehabilitation can be accomplished with a multidisciplinary approach, involving physiotherapist, play therapist and a child psychiatrist, in addition to the surgeon and primary care providers.

Control of hair growth using long-pulsed alexandrite laser is an efficient and cost effective therapy for patients suffering from recurrent pilonidal disease.

Pilonidal sinus (PNS) and its surgical management have a profound impact on hospital resources in terms of finances and productive man-hours. Surgical treatment has been the mainstay of treatment of both acute and chronic pilonidal sinus but recurrence is common. The control of hair growth in the sinus region plays an important role in preventing recurrence. Here, we discuss our experience of treating 19 patients suffering from recurrent pilonidal sinus with laser depilation and its long-term cost effectiveness. This is a retrospective study on patients who had recurrence of pilonidal sinus following multiple surgical treatments. They were treated using long-pulsed alexandrite laser for depilation in the sinus area, an outpatient procedure. Their clinical characteristics and outcomes were then evaluated. There was a significant reduction in hair density after laser treatment (p < 0.001). The disease-free period after laser treatment was significantly longer than that one after surgical treatment (p < 0.001). The average cost of repeated surgical treatment per disease-free month was significantly higher than that of laser treatment (p < 0.001). Evidence suggests the role of natal cleft hair growth in the evolution of the pilonidal disease; therefore, control of hair growth should be considered as an adjunct to the initial treatment via surgery. Compared to surgical treatment of recurrences, laser depilation is an efficient and cost-effective method of preventing recurrence and reducing morbidity and loss of man-hours. We suggest that laser depilation of the pilonidal sinus should be funded by clinical commissioning groups.

Utilization of genes encoding osmoprotectants in transgenic plants for enhanced abiotic stress tolerance

Global agriculture in the context of growing and expanding populations is under huge pressure to provide increased food, feed, and fiber. The recent phenomenon of climate change has further added fuel to the fire. It has been practically established now that the global temperature has been on the increase with associated fluctuations in annual rainfall regimes, and the resultant drought and flood events and increasing soil and water salinization. These challenges would be met with the introduction and utilization of new technologies coupled with conventional approaches. In recent years, transgenic technology has been proved very effective in terms of production of improved varieties of crop plants, resistant to biotic stresses. The abiotic stresses such as salt and drought are more complex traits, controlled by many genes. Transgenic plant development for these stresses has utilized many single genes. However, much emphasis has been placed on genes catalyzing the biosynthetic pathways of osmoprotectants. This review focuses on the current status of research on osmoprotectant genes and their role in abiotic stress tolerance in transgenic plants.

Common carotid artery dissection: a rare cause of acute neck swelling.

Spontaneous carotid artery dissection is a rare condition with potentially devastating consequences. Internal carotid artery and vertebral artery dissections have been implicated as the cause of 20% of strokes occurring in patients younger than 45 years. We describe a very rare case of a nontraumatic common carotid artery dissection in a 45-year-old man that was initially misdiagnosed as a sternocleidomastoid hematoma. This case highlights the need for vigilance for this often-missed diagnosis, as well as the indication for noninvasive imaging in unidentified neck swellings.

Surgical treatment of primary hyperparathyroidism: description of techniques and advances in the field.

Primary hyperparathyroidism is a disease commonly seen in patients above 60 years of age. It is the most common cause of asymptomatic or symptomatic hypercalcemia, usually found incidentally on routine check-ups. Surgical treatment is the only definitive treatment of choice in the symptomatic patient; however, it can also be employed in asymptomatic patients. First described in 1925, bilateral neck exploration is the gold standard of treatment for primary hyperparathyroidism. The recent interest in minimally invasive surgeries has led to better and improved techniques of neck exploration with improved cosmetic results and lesser chances of transient or permanent hypoparathyroidism due to inadvertent removal of normally functioning parathyroid tissue. These include unilateral neck explorations, minimally invasive parathyroidectomies and minimally invasive radio-guided parathyroidectomy. The intact parathyroid hormone assays have greatly added to the detection of normal and abnormal functioning glands, hence better surgical outcomes.