The pAblo·pCasso self-curing vector toolset for unconstrained cytidine and adenine base-editing in Gram-negative bacteria.

A synthetic biology toolkit, exploiting clustered regularly interspaced short palindromic repeats (CRISPR) and modified CRISPR-associated protein (Cas) base-editors, was developed for genome engineering in Gram-negative bacteria. Both a cytidine base-editor (CBE) and an adenine base-editor (ABE) have been optimized for precise single-nucleotide modification of plasmid and genome targets. CBE comprises a cytidine deaminase conjugated to a Cas9 nickase from Streptococcus pyogenes (SpnCas9), resulting in C→T (or G→A) substitutions. Conversely, ABE consists of an adenine deaminase fused to SpnCas9 for A→G (or T→C) editing. Several nucleotide substitutions were achieved using these plasmid-borne base-editing systems and a novel protospacer adjacent motif (PAM)-relaxed SpnCas9 (SpRY) variant. Base-editing was validated in Pseudomonas putida and other Gram-negative bacteria by inserting premature STOP codons into target genes, thereby inactivating both fluorescent proteins and metabolic (antibiotic-resistance) functions. The functional knockouts obtained by engineering STOP codons via CBE were reverted to the wild-type genotype using ABE. Additionally, a series of induction-responsive vectors was developed to facilitate the curing of the base-editing platform in a single cultivation step, simplifying complex strain engineering programs without relying on homologous recombination and yielding plasmid-free, modified bacterial cells.

Efecto del uso preoperatorio de bevacizumab intravítreo en pacientes con retinopatía diabética proliferativa sometidos a vitrectomía- Primera actualización / Preoperative intravitreal bevacizumab for proliferative diabetic retinopathy patients undergoing vitrectomy - First update

ACTUALIZACIÓN: Este resumen Epistemonikos (Living FRISBEE: Living FRIendly Summary of the Body of Evidence using Epistemonikos) es una actualización del resumen publicado en diciembre de 2014. INTRODUCCIÓN: La retinopatía diabética proliferativa conlleva un alto riesgo de ceguera si no es tratada de manera oportuna. El tratamiento muchas veces incluye la vitrectomía. El uso preoperatorio de bevacizumab, un anti-factor de crecimiento endotelial vascular, podría mejorar variables intraoperatorias que facilitan la cirugía y mejorarían el curso postoperatorio. MÉTODOS: Realizamos una búsqueda en Epistemonikos, la mayor base de datos de revisiones sistemáticas en salud, la cual es mantenida mediante el cribado de múltiples fuentes de información, incluyendo MEDLINE, EMBASE, Cochrane, entre otras. Extrajimos los datos desde las revisiones identificadas, analizamos los datos de los estudios primarios, realizamos un metanálisis y preparamos una tabla de resumen de los resultados utilizando el método GRADE. RESULTADOS Y CONCLUSIONES: Identificamos cinco revisiones sistemáticas que en conjunto incluyeron 16 estudios primarios, de los cuales 14 corresponden a ensayos aleatorizados. Concluimos que el uso preoperatorio de bevacizumab disminuye la incidencia de hemorragia vítrea en el postoperatorio temprano y probablemente también en el postoperatorio tardío, pero no está claro su efecto sobre la agudeza visual. Además, probablemente disminuye el tiempo quirúrgico, podría disminuir la incidencia de roturas retinianas iatrogénicas, y si bien no está claro que disminuya la ocurrencia de sangrado intraoperatorio, podría disminuir la necesidad de uso de endodiatermia.

UPDATE: This Living FRISBEE (Living FRIendly Summary of the Body of Evidence using Epistemonikos) is an update of the summary published in December 2014. INTRODUCTION: Proliferative diabetic retinopathy can cause severe vision loss and even blindness if left untreated. Vitrectomy is often required in the treatment of more severe cases. Preoperative administration of bevacizumab, a humanized anti-vascular endothelial growth factor would improve intraoperative variables that facilitate surgery and improve postoperative course. METHODS: We searched in Epistemonikos, the largest database of systematic reviews in health, which is maintained by screening multiple information sources, including MEDLINE, EMBASE, Cochrane, among others. We extracted data from the systematic reviews, reanalyzed data of primary studies, conducted a meta-analysis and generated a summary of findings table using the GRADE approach. RESULTS AND CONCLUSIONS: We identified five systematic reviews including 16 studies overall, of which 14 were randomized trials. We concluded the preoperative use of intravitreal bevacizumab reduces the rate of vitreous hemorrhage in the early postoperative period, and probably also in the late postoperative period, but its effect on visual acuity is not clear. Furthermore, it probably decreases the surgical time and may decrease the incidence of iatrogenic retinal breaks. Although we are uncertain whether preoperative bevacizumab decreases intraoperative bleeding, it may reduce the need for endodiathermy.

Coronary artery grafting in infants.

BACKGROUND: Coronary artery bypass grafting (CABG) with cardiac vale repair is an uncommon surgery in infants. CABG is technically demanding in infants due to the small size not only of the coronary arteries but also the potential graft arteries. The short and long-term outcome of surgery is not known and thus has largely been avoided. RESULTS: We report the case histories of two infants in whom CABG was undertaken successfully as a life-saving measure. Case 1: This infant needed an arterial switch operation after which the right coronary artery (RCA) was stenosed resulting in low cardiac output. After the right internal mammary artery (RIMA) was used to anastamose the RCA, the hemodynamic status improved drastically. Case 2: This infant underwent surgical correction for Anomalous Left Coronary Artery from Pulmonary Artery (ALCAPA). Postoperatively, she was in low cardiac output. She was found to have an occluded left coronary artery and mitral regurgitation (MR). After she underwent left internal mammary artery (LIMA) to Left Anterior Descending (LAD) anastamosis and mitral valve repair, the clinical condition improved dramatically. CONCLUSION: CABG is an uncommon operation in infants. This surgery is technically difficult. The long term results are not known and there are very few reports for the same. Though such an operation is best avoided, it can be used as a desperate life saving measure.

Successful use of blunt microdissection catheter in a chronic total occlusion of a celiomesenteric artery.

Chronic mesenteric ischemia is often a disabling condition associated with intestinal angina, weight loss, and sitophobia (a morbid aversion of food). Significant stenosis of two of the three main arteries of the gut is usually required to produce symptoms. Surgical therapy has included reimplantation and bypass grafting, usually with synthetic conduits and occasionally endarterectomy. Newer techniques have made endovascular treatment an emerging modality in managing some of the difficult lesions in the mesenteric circulation that cause chronic mesenteric ischemia. We describe the first reported case of blunt microdissection using a Frontrunner XP(R) CTO Catheter (Lumend, Redwood City, CA) to successfully cross, subsequently wire and stent a four-year-old chronic total occlusions in a celiac trunk, which also gave origin to the superior mesenteric artery.

First-trimester pregnancy loss and active Chlamydia trachomatis infection: correlation and ultrastructural evidence.

The incidence of Chlamydia trachomatis (Ct) infection and the possible correlation between couples presenting with first-trimester spontaneous abortions and active Ct infection was assessed. Additionally, the ability of Ct to infect zona-free hamster oocytes was explored by incubating the oocytes with spermatozoa from infected patients. A total of 961 women and 750 men consulting our reproductive medicine centre were screened for Ct using direct immunofluorescence. The general incidence of Ct infection was 9.4% in females (90 of 961) and 13.9% in males (104 of 750). In women with spontaneous abortions the incidence of Ct was 21.0% (14 of 66) compared with 8.9% (23 of 59) for women without spontaneous abortions and term pregnancies (chi-square, P < 0.05). When both partners of the couples were considered (one or both partners infected), the incidence rose to 68.8% (22 of 32) (chi-square, P < 0.001). In vitro studies using electron microscopy demonstrated the presence of Ct on the surface of and inside the oocyte. These results indicate a correlation between an active Ct infection and spontaneous abortion. Electron microscopy studies suggested the possibility of direct oocyte infection by Ct. Two models are proposed for the pathogenesis of Ct-related early abortions: (i) direct zygote infection, and (ii) immune response to heat shock proteins expressed by the zygote and triggered by previous Ct infections.

Damage control surgery.

The triad of hypothermia, acidosis, and coagulopathy during initial operative and resuscitation efforts has been recognized as a significant cause of death in patients with traumatic injuries. A staged surgical approach with a brief initial laparotomy, subsequent intensive care unit resuscitation, and a planned reoperation is an emerging technique used in trauma surgery, with application to a variety of other surgical challenges. Successful damage control therapy requires a coordinated multidisciplinary team effort by a trauma team experienced in the process of damage control operations, intensive care unit priorities, and potential complications of this innovative surgical approach.

[Exploratory analysis of risk factors of acute inner ear dysfunctions]. / Explorative Analyse von Risikofaktoren akuter Innenohrfunktionsstörungen.

Aside from the clinical picture, the frequent occurrence of vascular risk factors favors a vascular pathogenesis of acute sensorineural hearing loss. However, the data on the association of vascular risk factors with sensorineural hearing loss are controversial. Achieving a sufficient number of patients and establishing a suitable control group is difficult. We analyzed the data of 393 patients suffering from acute sensorineural hearing loss. Risk factors, general, audiological, and laboratory parameters were investigated. Mean hearing loss and remission during 10 days of hospital treatment were calculated. The patient group was divided into patients with a certain risk factor and those without this risk factor, and mean remission and hearing loss were compared. Patients suffering from hyperlipidemia or hypotension had substantial and significantly higher mean hearing losses than patients not suffering from these diseases. Thrombosis, embolism, and hypertension were associated with a significantly worse remission of hearing loss. Repeated episodes of sensorineural hearing loss showed a significant worse remission, but less mean hearing loss before therapy. Smoking, diabetes mellitus, infections of the upper airways, and allergies had no influence on remission and mean hearing loss.

Drawing with the non-dominant hand: implications for the study of construction.

BACKGROUND: Constructional impairment following left vs. right hemisphere damage has been extensively studied drawing tasks. A confounding factor in these studies is that right-handed patients with left hemisphere damage (LHD) are often forced by weakness to use their non-dominant (left) hand or hemiparetic dominant hand. Qualitative differences in the drawing characteristics of left and right hand drawings by normal subjects have not previously been characterized. The present study was undertaken to determine the qualitative differences between left and right hand drawings of normal subjects. METHODS: Thirty right-handed, elderly subjects without a history of neurological disease were asked to draw, from memory, seven objects using the right and left hand. Half of the subjects were randomly assigned to draw with the left hand first, and half the right hand first. Right and left hand drawings were compared using a standardized scoring system utilized in several previous studies of drawing in focal and diffuse neurological disease. Each drawing was scored on eighteen criteria. Right and left hand drawing scores were then compared using the t-test for paired samples or the Wilcoxon matched-pairs test. RESULTS: Drawings made using the left hand were found to be significantly simpler, more tremulous and of poorer overall quality than drawings made by the same subjects using the right hand. CONCLUSIONS: The deficits found in left versus right hand drawings of normals are similar to those found in patients with LHD, suggesting that much of the drawing impairment seen following LDH is due to an elementary motor disturbance related to use of the non-dominant hand.

Systemic lupus erythematosus: perinatal and neonatal implications.

Systemic lupus erythematosus (SLE) is a chronic autoimmune disorder that can affect almost all organ systems in the body. It is most common in women of childbearing age and may cause multiple peripartum complications. This article reviews the pathophysiology of SLE and the effects of SLE on fertility and pregnancy. The complexities of managing a pregnant patient with SLE are reviewed, and the importance of interdisciplinary collaboration discussed, as well as the effects of SLE on the fetus and a review of neonatal lupus erythematosus. Finally, a case report of a pregnant patient with SLE with challenging clinical management issues is presented.

Development of an alcohol withdrawal clinical pathway: an interdisciplinary process.

Studies show that as many as 40 percent of all patients in general hospitals are admitted because of complications related to alcoholism. However, the literature has little in specific clinical pathways, treatment protocols, or guidelines for the interdisciplinary care of these patients in the acute care setting. Furthermore, the little information that is published shows a lack of consistency in recommended treatment regimes. This article reviews a coordinated, interdisciplinary effort in developing and implementing a clinical pathway for alcohol withdrawal.

Serum S100beta release after coronary artery bypass grafting: roller versus centrifugal pump.

BACKGROUND: Microemboli generated during cardiopulmonary bypass (CPB) are implicated in the cerebral injury seen after coronary artery bypass grafting. Centrifugal pumps generate fewer microemboli than roller pumps. Increased S100beta levels have been reported after coronary artery bypass grafting, with levels greater than 1 ng/mL resulting in poorer neuropsychologic outcome. This study investigated the potential neurologic benefits of centrifugal pumps, by using S100beta as a marker for cerebral injury. METHODS: Thirty-two patients who had coronary artery bypass grafting were randomly assigned to two groups. Serial blood samples (preoperative, end of bypass, 30 minutes, and 2 and 24 hours after cardiopulmonary bypass) were taken and the serum analyzed for S100beta using a new immunoluminometric assay. RESULTS: Both groups were matched for age, number of grafts, and cardiopulmonary bypass and cross-clamp times. Postoperative serum S100beta levels were significantly higher in both groups than preoperative levels. Peak S100beta levels did not correlate with cardiopulmonary bypass time; however, 24-hour S100beta levels correlated with intubation time r = 0.40, p = 0.04). Th ere was no significant difference in S100beta levels between the groups at any of the time points. CONCLUSIONS: S100beta levels increased after coronary artery bypass grafting. Centrifugal pumps do not significantly decrease S100beta release. Persistently increased S100beta levels are associated with longer intubation times.

Integrating nursing education into a trauma outreach program.

A level 1 trauma center is challenged to provide optimal trauma care to patients in the region it serves. One approach is to increase the knowledge and clinical skills of all trauma care providers, especially those who work outside urban areas. This article describes one trauma center's experience of integrating rural nursing education into an existing medical trauma outreach program. The educational strategies, example of course content, and organizational aspects of the program are described, along with marketing strategies used to promote the program.

[Ultrasonography-guided transvaginal puncture of ovarian cysts]. / Punción vía transvaginal de quistes ováricos guiada por ultrasonografía.

Presenting the experience in 18 cases of ovarian cystic transvaginal puncture guided by ultrasonography. The cystic selected for puncture were unilateral, regular contours, uniloculated without internal papillae, without ascites. This procedure was made in women on reproductive age with failure to medical treatment.

Transdermal delivery of human insulin to albino rabbits using electrical current.

The transdermal route of administration for medication has many potential advantages over other routes of administration. However, the stratum corneum is an effective barrier to the absorption of most chemicals from the external environment into the body. To evaluate techniques for alteration of transdermal permeability, the authors studied the effect of low levels of electrical current on transport of a protein across the stratum corneum. Transcutaneous insulin absorption was used as an indicator of altered permeability. Twenty-six albino rabbits had acute diabetes mellitus induced by the intravenous administration of 125 mg/kg of alloxan. The animals then received either cutaneous patches containing insulin and an electrical current of 0.4 mA (active) or patches containing an equal amount of insulin but without electrical current (passive). At 10 and 12 hours after the placement of the patches, animals with active patches had significant elevations in serum insulin levels (p less than .05) and reduction in blood glucose levels (p less than .01). No changes were seen in controls. Animals with active patches also had significant differences from control animals in mean insulin response and peak insulin response (p less than .05). No cutaneous toxicity was observed in any of the animals. The authors conclude that low levels of electrical current can induce changes in stratum corneum permeability that are sufficient to produce the transdermal absorption of physiologic doses of a protein such as human insulin.

[Intravenous leiomyomatosis]. / Leiomiomatosis intravenosa.

A case of intravascular leiomiomatosis is presented. This gynecologic diseases is rare, although it can be of risk. A case is described with a review of the literature.

New insights on the mode of action of intrauterine contraceptive devices in women.

To gain a better understanding of the mechanism of action of intrauterine devices (IUDs), a search was made for ova in the genital tracts of 115 women using no contraception and of 56 women using IUDs, all of whom volunteered for study in conjunction with surgical sterilization. Ova were recovered from tubal flushings between 48 and 120 hours after the midcycle peak of luteinizing hormone in 39% of the IUD users compared with 56% of women in the control group (0.05 less than P less than 0.10). This suggests an action of the IUD before the ovum reaches the uterus. Eggs with a microscopic appearance consistent with fertilization were recovered from the fallopian tubes of half of the women using no contraception who had intercourse within the fertile period of the reproductive cycle and from whom ova were recovered. In contrast (P less than 0.01), no eggs with this appearance were recovered in IUD users who had intercourse within the fertile period. No ova were recovered from the body of the uterus of any of the IUD users. Fertilized ova are less likely to reach the uterine cavity containing an IUD. Thus, the principal mode of IUDs is by a method other than destruction of live embryos.

Treatment of leiomyomata uteri with D-Trp6-luteinizing hormone-releasing hormone.

Suppression of the secretion of gonadal steroids by chronic administration of a superactive agonist of luteinizing hormone-releasing hormone (LH-RH) was used for treatment of leiomyomata uteri. Ten menstruating women, presenting with a total of 20 uterine leiomyomas, were treated for 3 months with daily subcutaneous injections of D-Trp6-LH-RH. Serum estradiol (E2) levels were suppressed rapidly in five patients and were decreased in other patients. At the end of therapy, leiomyomas regressed completely in three patients, while five patients showed a decrease of more than 40% in the volume of leiomyomas. The reduction in tumor size was correlated with the rapidity of the fall in serum E2 levels. In one patient, the leiomyomata increased in size during treatment, and one woman had a poor clinical response. The agonist was well tolerated and few side effects were observed. Therapy with LH-RH agonists offers an alternative in the management of some uterine leiomyomas.