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Mean age in the hip fracture cohort was 80.9 years and in the combined fracture group 80.3 years. Out of the 40 patients in the combined group four were treated operatively. The 30-d mortality for our hip fracture cohort was 7.2% compared to the mortality of our combined cohort of 12.5% (

= 0.163). The one-year mortality for our hip fracture cohort was 26.4% compared to 40% for the combined fracture cohort (

= 0.038). We also found patients with combined injuries were less likely to return to their own home.

The 30-d and one-year mortality is higher for those patients who have sustained a combined hip and proximal humerus fracture when compared to those with a hip fracture alone.

The 30-d and one-year mortality is higher for those patients who have sustained a combined hip and proximal humerus fracture when compared to those with a hip fracture alone.

The incidence of primary osteoarthritis knee is gradually increasing among young individuals. The increasing prevalence of obesity, sedentary lifestyle, sporting activity, and vitamin D deficiency (VDD) has been hypothesized for this shifting disease trend. This study was designed to look for the association of serum vitamin D among these young arthritic patients.

To look for the association of serum vitamin D in younger knee osteoarthritis (KOA) patients.

In a 2-year observational study, 146 non-obese KOA patients of 35-60 years were evaluated clinically (Knee injury and Osteoarthritis Outcome Score, KOOS) and radiologically (Kellegren-Lawrence stage, KL). The serum 25(OH)D level of these patients and 146 normal healthy individuals of same age group were estimated.

Both the groups were comparable in terms of age and sex. The average serum 25(OH)D level in healthy individuals and KOA patients was 45.83 ng/mL and 34.58 ng/mL, respectively (

< 0.001). Inadequate serum 25(OH)D level (< 30 ng/mL) was found in 46.57% of KOA patients and 24% of normal healthy participants indicating a significant positive association (odds ratio 2.77, 95%CI 1.67-4.54,

< 0.001). The 25(OH)D level in KL grade I, II, III and IV was 43.40, 30.59, 31.56 and 33.93 ng/mL respectively (no difference,

= 0.47). Similarly, the KOOS score in sufficient, insufficient and deficient groups were 65.31, 60.36 and 65.31, respectively (no difference,

= 0.051).

The serum 25(OH)D level is significantly low in younger KOA patients. However, the clinical and radiological severities have no association with serum vitamin D level.

The serum 25(OH)D level is significantly low in younger KOA patients. However, the clinical and radiological severities have no association with serum vitamin D level.Therapeutic applications of enzymes have been widely accepted in clinical practices for decades. Proteolytic enzymes in particular, have been used for the treatment of diseases and disorders. Serratiopeptidase is a proteolytic enzyme having immense applications in therapeutic areas which have been validated by several in vitro, in vivo, and clinical studies as well as through anecdotal evidences. These applications are attributable to its versatile properties including anti-inflammatory, anti-biofilm, analgesic, anti-edemic, and fibrinolytic effects. The significant impact of serratiopeptidase reported needs to be backed by more scientific data. This review encompasses the details of therapeutic applications of serratiopeptidase based on available in vitro, in vivo, and clinical studies. We found some strong evidences regarding the efficacy of serratiopeptidase. However data on safety, tolerability, and its mechanism of action need detailing. This review aims to further explore the available literature on serratiopeptidase as well as provide scientific details for existing applications.Per- and polyfluoroalkyl substances (PFAS) are synthetic organic compounds that over the past several years, have witnessed a dramatic increase in scientific attention. As PFAS are predominantly accumulated in plasma, monitoring individual burden levels in plasma are typically achieved via some combination of protein precipitation and/or solid phase extraction (SPE), either in online or offline modes. This work describes an updated PFAS extraction workflow, using 96-well plate technology and protein precipitation that is rapid, simple, inexpensive, and amenable for large cohort studies. In brief, plasma proteins were precipitated using methanol and the resulting centrifuged supernatant was directly analyzed using UHPLC-MS/MS. We monitored 51 PFAS, which were quantified via isotope dilution and the effectiveness of the method was demonstrated by using NIST blood-based Standard Reference Materials (SRMs). This method resulted in recoveries ranging between 70 and 89% for all analytes. The 96-well design exhibited low limits of detection and only required sample volumes of 100 µL, thus resulting in an amenable method for high-throughput plasma/serum PFAS screening. • PFAS were directly quantified in plasma and serum samples; • No SPE needed after protein precipitation; • SRMs can be used to validate PFAS measurement in plasma/serum.

To describe the

bacterial adhesion protocol of

and

asei on dental surfaces for a qualitative approach by Scanning Electron Microscopy (SEM) observations. A control and Amelogenesis Imperfecta (AI) affected teeth were used to validate the protocol.

Eight teeth were collected and fixed in 10% formalin during 10 days. Crowns were fragmented into 4 parts and kept in the freshly prepared artificial saliva. For the preparation of bacterial suspensions, bacterial strains (

and L.

) were incubated in a freshly prepared culture medium. After two successive cultures at 37°C and 3 rinces, bacterial suspensions were prepared in artificial saliva and adjusted to correspond to 10

CFU ml

. Bacterial adhesion was carried out by sedimentation. Dental fragments were immersed in bacterial suspensions and rinsed with PBS to remove non adherent bacteria. Adherent bacteria were fixed with glutaraldehyde. Finally, teeth samples were dehydrated, coated, dried and observed using high-resolution SEM (JEOL, JSM-5400).

SEM observations showed adherence of spheric stuctures, identified as

and bacilic structures identified as L.

.

Adhesion of bacteria could be observed by SEM and depends on the quality of dental mineralized tissues.

Adhesion of bacteria could be observed by SEM and depends on the quality of dental mineralized tissues.CRISPR/Cas systems (Clustered regularly interspaced palindromic repeats / CRISPR-associated) are rapidly becoming a commonplace and popular tool for gene editing in research and clinical contexts. However, the quality of CRISPR/Cas experiments depends heavily on the guide RNA (gRNA) design; therefore, a reliable, easy, and rapid method for verifying gRNA cleavage efficacy is necessary. Engineered nuclease-induced translocations (ENIT) are an easy and cost-efficient method for the verification of gRNA efficacy, which involves tracking induced chromosomal mutations, using polymerase chain reaction (PCR). We have customized this method using both direct PCR and nested PCR approaches and have been able to reduce the sample preparation time. We present a simple and reliable gRNA testing approach that requires no specific enzymes or equipment.•The approach requires only routinely used enzymes and equipment.•Cost- and time-efficient, requiring approximately 30 min for PCR sample preparation, without requiring DNA purification.•High sensitivity, with induced translocation detected in 100 of 10,000 cells in the general population.Baculovirus expression vector systems (BEVS) have been widely used for production of recombinant proteins in insect cells. However, baculoviruses superinfection and repeated passages originate defective interfering particle (DIP) mutants, which is a limitation to a continuous large-scale production. Accordingly, a classical chemical transfection method performed on monolayer of Spodoptera frugiperda insect cells (Sf9) was modified to produce recombinant baculoviruses with high efficiency. Modifications consist to transfect exponentially growing cells in suspension after concentration by tenfold through centrifugation. Ten different constructions of recombinant baculoviruses with insert varying in size from 180 bp to 2,395 bp, were obtained through employment of the Bac-to-Bac expression system (ThermoFisher/Invitrogen). The transfection efficiency of the modified protocol varied from 45 to 57%, independent of the insert size, while classical method present transfection efficiency of 2 to 20%. EGFR tumor After transfection of 6 × 106 cells, the recombinant baculoviruses titer obtained with modified method was about 2 × 107 pfu/ mL in a total volume of 12 mL, which is scalable to 24 liters of 1 × 108 pfu/ mL, after only two amplification rounds, contributing to improve large scale heterologous protein production in insect cells, with low amplification passages.Primary human vulvovaginal fibroblast cell lines are useful for studying biological mechanisms underlying genital pain, pelvic organ prolapse, and the spread of sexually transmitted infections. However, the vaginal biopsies necessary for establishing these cell lines are invasive and relatively difficult to obtain. Primary mouse fibroblast cell lines derived from pre-clinical animal models of these conditions can be used for better controlled experiments that can help us dissect disease mechanisms. To our knowledge, there are no published protocols for establishing primary murine vaginal fibroblast cell lines to date. Here, we describe a protocol for the establishment of murine vaginal fibroblast cell lines via enzymatic digestion of vaginal canal tissue. Cell lines generated using this method can be used for in vitro studies of these important structural cells in a variety of pre-clinical mouse models; such studies are required to identify and characterize relevant regulatory and therapeutic targets in a wide array of diseases of interest. As shown in our representative data, this protocol yields viable cell lines from ND4 Swiss outbred mice. These cells bear surface markers characteristic of fibroblasts and are capable of producing inflammatory cytokines in response to treatment with bacterial and yeast antigens in vitro.This study aimed to introduce a new method for eye lens thermo-luminescent dosimetry and also estimate the dose associated with induced cancer risk due to the ionizing radiation exposure received by physicians and other staff cooperating in interventional cardiology (IC) procedures. The measurements were performed with six TLDs (thermoluminescent dosimeters) four TLDs for eye lens dosimetry (2 positioned on respiratory/surgical mask under the eye region as the new method; and 2 near the outside border of the eye as the common method) and two TLDs for whole-body dosimetry. Whole-body doses were used to calculate the cancer risks induced by IC procedures. The results of the new proposed method for eye lens dosimetry were similar to common TLD positioning (mean differences less then 5%) and mask displacement had no significant effect on eye dose measurement in our new method. Our proposed method for eye lens dosimetry is simpler and more comfortable compared to the common method and it can be used as an alternative method without using TLD holders to monitor lens dose for IC workers wearing masks during the procedure.