The Surgical Site Infection Risk Score (SSIRS): A Model to Predict the Risk of Surgical Site Infections

Background

Surgical site infections (SSI) are an important cause of peri-surgical morbidity with risks that vary extensively between patients and surgeries. Quantifying SSI risk would help identify candidates most likely to benefit from interventions to decrease the risk of SSI.

Methods

We randomly divided all surgeries recorded in the National Surgical Quality Improvement Program from 2010 into a derivation and validation population. We used multivariate logistic regression to determine the independent association of patient and surgical covariates with the risk of any SSI (including superficial, deep, and organ space SSI) within 30 days of surgery. To capture factors particular to specific surgeries, we developed a surgical risk score specific to all surgeries having a common first 3 numbers of their CPT code.

Results

Derivation (n = 181 894) and validation (n = 181 146) patients were similar for all demographics, past medical history, and surgical factors. Overall SSI risk was 3.9%. The SSI Risk Score (SSIRS) found that risk increased with patient factors (smoking, increased body mass index), certain comorbidities (peripheral vascular disease, metastatic cancer, chronic steroid use, recent sepsis), and operative characteristics (surgical urgency; increased ASA class; longer operation duration; infected wounds; general anaesthesia; performance of more than one procedure; and CPT score). In the validation population, the SSIRS had good discrimination (c-statistic 0.800, 95% CI 0.795–0.805) and calibration.

Conclusion

SSIRS can be calculated using patient and surgery information to estimate individual risk of SSI for a broad range of surgery types.     

Qualichem In Vivo: A Tool for Assessing the Quality of In Vivo Studies and Its Application for Bisphenol A

In regulatory toxicology, quality assessment of in vivo studies is a critical step for assessing chemical risks. It is crucial for preserving public health studies that are considered suitable for regulating chemicals are robust. Current procedures for conducting quality assessments in safety agencies are not structured, clear or consistent. This leaves room for criticism about lack of transparency, subjective influence and the potential for insufficient protection provided by resulting safety standards. We propose a tool called “Qualichem in vivo” that is designed to systematically and transparently assess the quality of in vivo studies used in chemical health risk assessment. We demonstrate its use here with 12 experts, using two controversial studies on Bisphenol A (BPA) that played an important role in BPA regulation in Europe. The results obtained with Qualichem contradict the quality assessments conducted by expert committees in safety agencies for both of these studies. Furthermore, they show that reliance on standardized guidelines to ensure scientific quality is only partially justified. Qualichem allows experts with different disciplinary backgrounds and professional experiences to express their individual and sometimes divergent views—an improvement over the current way of dealing with minority opinions. It provides a transparent framework for expressing an aggregated, multi-expert level of confidence in a study, and allows a simple graphical representation of how well the study integrates the best available scientific knowledge. Qualichem can be used to compare assessments of the same study by different health agencies, increasing transparency and trust in the work of expert committees. In addition, it may be used in systematic evaluation of in vivo studies submitted by industry in the dossiers that are required for compliance with the REACH Regulation. Qualichem provides a balanced, common framework for assessing the quality of studies that may or may not be following standardized guidelines.     

Phagocytic Cells in the Peripheral Blood of the Dogfish Scyliorhinus canicula L. I. In Vitro Studies

In vitro phagocytosis by peripheral blood leucocytes of the dogfish Scyliorhinus canicula L. was examined by exposing adherent cells to a variety of particulate and soluble antigens and inert material. Their subsequent uptake was monitored by light, scanning and transmission electron microscopy. The monocyte and the neutrophil-like granulocyte were found to be the major phagocytic cells. Larger particles like yeasts and erythrocytes were the most avidly phagocytosed. From studies on the effects of pH, temperature and the presence of plasma, metabolic inhibitors and divalent ions, it appeared that optimum phagocytosis occurred at pH 7.0 and between 10 and 20°C. Serum factors did not enhance the process in this species. Finally, the in vitro clearance of 5 bacterial species indicated that the presence of blood phagocytes had little or no effect on bacterial numbers.     

Specific Role of Each Human Leukocyte Type in Viral Infections I. Monocyte as Host Cell for Vesicular Stomatitis Virus Replication In Vitro

Each major leukocyte type of the peripheral blood of healthy donors was studied in vitro for its ability to support vesicular stomatitis virus (VSV) replication. Purified cultures of each white blood cell type were prepared by the selective adsorption and elution of cells from silicone-treated glass beads. It was found that monocytes and macrophages (derived from the rapid transformation of monocytes in vitro) were the principal host cells for VSV replication. Interferon added to mixed leukocyte cultures, prior to virus inoculation, reduced virus yields and prevented destruction of macrophages. Cultures of small lymphocytes, containing no detectable monocytes or macrophages, produced amounts of virus equivalent to 1% of that produced in leukocyte cultures which contained 7% monocytes. Small lymphocytes did not undergo demonstrable cytopathic alterations in virus-infected cultures. VSV neither replicated nor produced cytopathic effects in polymorphonuclear leukocytes.     

Murine Mammary Tumour Cells In Vitro. I. the Development of A Quiescent State

Abstract Three mouse mammary tumour lines (66, 67, and 68H) derived from a single mouse mammary tumour were investigated for their growth kinetics and development of quiescent cells in unfed monolayer cultures. All three lines develop pure quiescent populations when grown in unfed plateau cultures. A dramatic cell-cycle redistribution accompanied the proliferating (P) to quiescent (Q) transition, with the percentage of cells having a G₁ DNA content increasing from 50% in the P state to <97% in the Q state. As the cultures progressed from exponential to plateau growth, a decrease of 50% in cellular RNA was observed in all three lines. This property enables the clear identification of P v. Q cells by flow cytometry using the two-step acridine orange assay. Autoradiographic data verified that these plateau cells were quiescent since >2.5% of the cells incorporated [³H]TdR when labelled for approximately two doubling times. Further comparison of the P and Q cells showed that: (a) the Coulter volume of Q cells was approximately half that of P cells in all three lines; (b) viability, as measured by dye exclusion was >95% in all cultures regardless of their proliferative state; and (c) colony-forming ability decreased as the cells entered the quiescent state. In each of these cell lines the development of Q-cell populations was marked by similar changes in all measured parameters. These quiescent tumour cells provide a relatively simple model to evaluate what, if any, important differences exist between the response of P v. Q cells to various therapeutic agents.     

Modelling Neuroinflammation In Vitro: A Tool to Test the Potential Neuroprotective Effect of Anti-Inflammatory Agents

Neuron-microglia co-cultures treated with pro-inflammatory agents are a useful tool to study neuroinflammation in vitro, where to test the potential neuroprotective effect of anti-inflammatory compounds. However, a great diversity of experimental conditions can be found in the literature, making difficult to select the working conditions when considering this approach for the first time. We compared the use of neuron-primary microglia and neuron-BV2 cells (a microglial cell line) co-cultures, using different neuron:microglia ratios, treatments and time post-treatment to induce glial activation and derived neurotoxicity. We show that each model requires different experimental conditions, but that both neuron-BV2 and neuron-primary microglia LPS/IFN-γ-treated co-cultures are good to study the potential neuroprotective effect of anti-inflammatory agents. The contribution of different pro-inflammatory parameters in the neurotoxicity induced by reactive microglial cells was determined. IL-10 pre-treatment completely inhibited LPS/IFN-γ-induced TNF-α and IL-6 release, and COX-2 expression both in BV2 and primary microglial cultures, but not NO production and iNOS expression. However, LPS/IFN-γ induced neurotoxicity was not inhibited in IL-10 pre-treated co-cultures. The inhibition of NO production using the specific iNOS inhibitor 1400 W totally abolished the neurotoxic effect of LPS/IFN-γ, suggesting a major role for NO in the neurotoxic effect of activated microglia. Consequently, among the anti-inflammatory agents, special attention should be paid to compounds that inhibit NO production.     

Co-Culture of S. epidermidis and Human Osteoblasts on Implant Surfaces: An Advanced In Vitro Model for Implant-Associated Infections

Objectives

Total joint arthroplasty is one of the most frequent and effective surgeries today. However, despite improved surgical techniques, a significant number of implant-associated infections still occur. Suitable in vitro models are needed to test potential approaches to prevent infection. In the present study, we aimed to establish an in vitro co-culture setup of human primary osteoblasts and S. epidermidis to model the onset of implant-associated infections, and to analyze antimicrobial implant surfaces and coatings.

Materials and Methods

For initial surface adhesion, human primary osteoblasts (hOB) were grown for 24 hours on test sample discs made of polystyrene, titanium alloy Ti6Al4V, bone cement PALACOS R^®, and PALACOS R^® loaded with antibiotics. Co-cultures were performed as a single-species infection on the osteoblasts with S. epidermidis (multiplicity of infection of 0.04), and were incubated for 2 and 7 days under aerobic conditions. Planktonic S. epidermidis was quantified by centrifugation and determination of colony-forming units (CFU). The quantification of biofilm-bound S. epidermidis on the test samples was performed by sonication and CFU counting. Quantification of adherent and vital primary osteoblasts on the test samples was performed by trypan-blue staining and counting. Scanning electron microscopy was used for evaluation of topography and composition of the species on the sample surfaces.

Results

After 2 days, we observed approximately 10⁴ CFU/ml biofilm-bound S. epidermidis (10³ CFU/ml initial population) on the antibiotics-loaded bone cement samples in the presence of hOB, while no bacteria were detected without hOB. No biofilm-bound bacteria were detectable after 7 days in either case. Similar levels of planktonic bacteria were observed on day 2 with and without hOB. After 7 days, about 10⁵ CFU/ml planktonic bacteria were present, but only in the absence of hOB. Further, no bacteria were observed within the biofilm, while the number of hOB was decreased to 10% of its initial value compared to 150% in the mono-culture of hOB.

Conclusion

We developed a co-culture setup that serves as a more comprehensive in vitro model for the onset of implant-associated infections and provides a test method for antimicrobial implant materials and coatings. We demonstrate that observations can be made that are unavailable from mono-culture experiments.     

In Vitro Studies of the Metabolism of Tissue Slices : I. Metabolic and Physical Factors Influencing the Penetration of Tetrazolium Salts

The use of tetrazolium salts for metabolic studies has been dismissed on the basis of their poor penetration into fresh tissue slices. In view of the fact that the penetration of these compounds can be visualized, it was felt that knowledge of the factors involved would be important. Factors, known to influence the penetration of oxygen, were examined with respect to the tetrazolium salts. The penetration of tetrazolium salt into tissue slices was a regular and predictable phenomenon. It was found that decreasing the metabolism of the cells in the slice substantially increased the penetration of these compounds, while increases in metabolism, by addition of substrate (such as succinate) to the incubating medium, considerably decreased their penetration. Increasing concentrations of the salt in the medium resulted in greater but limited penetration. It is our belief that the metabolism of tissue slices can be effectively studied with the aid of the tetrazolium salts, the portion of the population of cells participating in any reaction being accurately established by measuring the depth of the zone of reduced dye.     

Cephalothin, a New Cephalosporin with a Broad Antibacterial Spectrum: I. In Vitro Studies Employing the Gradient Plate Technique

Cephalothin is 7-(thiophene-2-acetamido) cephalosporanic acid; it was prepared by N-acylation of the nucleus of cephalosporin C, 7-aminocephalosporanic acid. Cephalothin had a broad spectrum of antibiotic activity that was essentially unaffected by human serum or inoculum level, the activity of penicillinase, or pH variation of the growth medium. In vitro development of resistance by staphylococci could not be demonstrated, but the gram-negative organisms did develop a stepwise type of resistance to the antibiotic. Staphylococci made resistant in vitro to 5-methyl-3-phenyl-4-isoxazole penicillin were also resistant to cephalothin and to 6-(2,6-dimethoxybenzamido) penicillin; however, the mechanism of resistance to each antibiotic may have differed. Some complications involved in the laboratory evaluation methods currently in use in the field of antibiotics are examined.     

Enzymes of Krebs-Henseleit Cycle in Vitis vinifera L: III. In Vivo and In Vitro Studies of Arginase

Ornithine carbamoyltransferase (OCT) activity was detected in extracts from mature leaves, fruit, germinating seeds, and seedlings of Vitis vinifera L. Michaelis-Menten constants for OCT were 3.5 millimolar for carbamyl phosphate and 5.5 millimolar for l-ornithine. Concentrations of l-ornithine greater than 10 millimolar slightly inhibited the enzyme, whereas carbamyl phosphate at concentrations greater than the optimal (about 10 millimolar) did not affect OCT activity. l-Citrulline formation was linear with incubation period for the first 25 minutes and with increasing amounts of enzyme up to an equivalent of about 200 milligrams of fresh tissue. The optimum pH for in vitro OCT activity was between 8.4 and 8.8, and the optimum incubation temperature was 38 C.     

Atomic Force Microscopy: A Promising Tool for Deciphering the Pathogenic Mechanisms of Fungi in Cystic Fibrosis

During the past decades, atomic force microscopy (AFM) has emerged as a powerful tool in microbiology. Although most of the works concerned bacteria, AFM also permitted major breakthroughs in the understanding of physiology and pathogenic mechanisms of some fungal species associated with cystic fibrosis. Complementary to electron microscopies, AFM offers unprecedented insights to visualize the cell wall architecture and components through three-dimensional imaging with nanometer resolution and to follow their dynamic changes during cell growth and division or following the exposure to drugs and chemicals. Besides imaging, force spectroscopy with piconewton sensitivity provides a direct means to decipher the forces governing cell–cell and cell–substrate interactions, but also to quantify specific and non-specific interactions between cell surface components at the single-molecule level. This nanotool explores new ways for a better understanding of the structures and functions of the cell surface components and therefore may be useful to elucidate the role of these components in the host–pathogen interactions as well as in the complex interplay between bacteria and fungi in the lung microbiome.

     

PHISDetector: A Tool to Detect Diverse In Silico Phage–host Interaction Signals for Virome Studies

Phage–microbe interactions are appealing systems to study coevolution, and have also been increasingly emphasized due to their roles in human health, disease, and the development of novel therapeutics. Phage–microbe interactions leave diverse signals in bacterial and phage genomic sequences, defined as phage–host interaction signals (PHISs), which include clustered regularly interspaced short palindromic repeats (CRISPR) targeting, prophage, and protein–protein interaction signals. In the present study, we developed a novel tool phage–host interaction signal detector (PHISDetector) to predict phage–host interactions by detecting and integrating diverse in silico PHISs, and scoring the probability of phage–host interactions using machine learning models based on PHIS features. We evaluated the performance of PHISDetector on multiple benchmark datasets and application cases. When tested on a dataset of 758 annotated phage–host pairs, PHISDetector yields the prediction accuracies of 0.51 and 0.73 at the species and genus levels, respectively, outperforming other phage–host prediction tools. When applied to on 125,842 metagenomic viral contigs (mVCs) derived from 3042 geographically diverse samples, a detection rate of 54.54% could be achieved. Furthermore, PHISDetector could predict infecting phages for 85.6% of 368 multidrug-resistant (MDR) bacteria and 30% of 454 human gut bacteria obtained from the National Institutes of Health (NIH) Human Microbiome Project (HMP). The PHISDetector can be run either as a web server (http://www.microbiome-bigdata.com/PHISDetector/) for general users to study individual inputs or as a stand-alone version (https://github.com/HIT-ImmunologyLab/PHISDetector) to process massive phage contigs from virome studies.     

In Vitro Efficient Transfection by CM18-Tat11 Hybrid Peptide: A New Tool for Gene-Delivery Applications

Cell penetrating peptides (CPPs) are actively researched as non-viral molecular carriers for the controlled delivery of nucleic acids into cells, but widespread application is severely hampered by their trapping into endosomes. Here we show that the recently introduced endosomolytic CM₁₈-Tat₁₁ hybrid peptide (KWKLFKKIGAVLKVLTTG-YGRKKRRQRRR, residues 1-7 of Cecropin-A, 2-12 of Melittin, and 47-57 of HIV-1 Tat protein) can be exploited to obtain a self-assembled peptide-DNA vector which maintains the CM₁₈-Tat₁₁ ability to enter cells and destabilize vesicular membranes, concomitantly yielding high DNA transfection efficiency with no detectable cytotoxic effects. Different peptide-DNA stoichiometric ratios were tested to optimize vector size, charge, and stability characteristics. The transfection efficiency of selected candidates is quantitatively investigated by the luciferase-reporter assay. Vector intracellular trafficking is monitored in real time and in live cells by confocal microscopy. In particular, fluorescence resonant energy transfer (FRET) between suitably-labeled peptide and DNA modules was exploited to monitor complex disassembly during endocytosis, and this process is correlated to transfection timing and efficiency. We argue that these results can open the way to the rational design and application of CM₁₈-Tat₁₁–based systems for gene-delivery purposes.     

水稻高脯氨酸愈伤组织变异体的选择及其耐盐性

以水稻（ＯｒｙｚａｓａｔｉｖａＬ．）品种双丰１号幼胚愈伤组织为材料,经５０ＣＯγ－射线诱变处理后,用羟基脯氨酸（Ｈｙｐ）作为选择压力,通过多次在含Ｈｙｐ（３．０ｍｍｏｌ／Ｌ）的选择培养基和无Ｈｙｐ培养基上交替培养,分离出耐Ｈｙｐ的愈伤组织变异体。该变异体以高脯氨酸含量为特征,其脯氨酸含量比原型高２．６倍;在ＮａＣｌ处理时,随着ＮａＣｌ浓度的提高,变异体愈伤组织脯氨酸含量的增加大大高于原型。这种高脯氨酸变异体的耐盐性较原型强,用相对鲜重估计其耐性比对照提高３９％。变异体在盐胁迫下可溶性糖含量比原型多,吸收较多的Ｎａ＋,并维持较高的Ｋ＋含量;但Ｋ＋／Ｎａ＋比与原型无差异。变异体含水量略有下降。这些性状的变化显示了变异体在盐胁迫下渗透调节的加强。     

Release of Metal Ions from Orthodontic Appliances by In Vitro Studies: A Systematic Literature Review

In the present work, a systematic literature review on release of metal ions from orthodontic appliances under in vitro conditions is described. Detailed and schematic analysis of used materials and applied methods (immersion media, incubation time, temperature, and analytical techniques) is provided. The PubMed search identified 40 studies, among which eight met the selection criteria. One additional study was included in the review. All the authors agreed that the doses of released metal ions were far below the toxic level and the dietary intake. Although the concentrations of metal ions in immersion media greatly differed, the general conclusions were coherent. It must be underlined that the main disadvantage of in vitro tests was that the experimental setup did not reflect in vivo conditions, e.g., the presence of biofilm, which grows on the surface of the materials in oral cavity. The presence and activity of microflora to a large extent is responsible for the process of corrosion, in particular, biodeterioration. The further scheme of in vitro research should incorporate changeable conditions of oral cavity environment (pH, dynamic conditions—saliva flow) and the presence of microbiological flora (microbiological attack) in the experimental design and, first of all, the real proportions of appliance elements.     

Dual Beneficial Effects of (-)-Epigallocatechin-3-Gallate on Levodopa Methylation and Hippocampal Neurodegeneration: In Vitro and In Vivo Studies

Background

A combination of levodopa (L-DOPA) and carbidopa is the most commonly-used treatment for symptom management in Parkinson''s disease. Studies have shown that concomitant use of a COMT inhibitor is highly beneficial in controlling the wearing-off phenomenon by improving L-DOPA bioavailability as well as brain entry. The present study sought to determine whether (-)-epigallocatechin-3-gallate (EGCG), a common tea polyphenol, can serve as a naturally-occurring COMT inhibitor that also possesses neuroprotective actions.

Methodology/Principal Findings

Using both in vitro and in vivo models, we investigated the modulating effects of EGCG on L-DOPA methylation as well as on chemically induced oxidative neuronal damage and degeneration. EGCG strongly inhibited human liver COMT-mediated O-methylation of L-DOPA in a concentration-dependent manner in vitro, with an average IC ₅₀ of 0.36 µM. Oral administration of EGCG moderately lowered the accumulation of 3-O-methyldopa in the plasma and striatum of rats treated with L-DOPA + carbidopa. In addition, EGCG also reduced glutamate-induced oxidative cytotoxicity in cultured HT22 mouse hippocampal neuronal cells through inactivation of the nuclear factor κB-signaling pathway. Under in vivo conditions, administration of EGCG exerted a strong protective effect against kainic acid-induced oxidative neuronal death in the hippocampus of rats.

Conclusions/Significance

These observations suggest that oral administration of EGCG may have significant beneficial effects in Parkinson''s patients treated with L-DOPA and carbidopa by exerting a modest inhibition of L-DOPA methylation plus a strong neuroprotection against oxidative damage and degeneration.     

Proportions of Staphylococcus aureus and Methicillin-Resistant Staphylococcus aureus in Patients with Surgical Site Infections in Mainland China: A Systematic Review and Meta-Analysis

Background

Sufficient details have not been specified for the epidemiological characteristics of Staphylococcus aureus (S. aureus) and methicillin-resistant Staphylococcus aureus (MRSA) among surgical site infections (SSIs) in mainland China. This systematic review aimed to estimate proportions of S. aureus and MRSA in SSIs through available published studies.

Methods

PubMed, Embase and four Chinese electronic databases were searched to identify relevant primary studies published between 2007 and 2012. Meta-analysis was conducted on the basis of logit-transformed metric for proportions of S. aureus and MRSA, followed by pre-defined subgroup meta-analysis. Random-effects meta-regression was also conducted to explore the impact of possible factors on S. aureus proportions.

Results

106 studies were included, of which 38 studies involved MRSA. S. aureus accounted for 19.1% (95%CI 17.2-21.0%; I² = 84.1%) of all isolates in SSIs, which was roughly parallel to 18.5% in the United States (US) (P-value = 0.57) but significantly exceeded those calculated through the surveillance system in China (P-value<0.001). In subgroup analysis, S. aureus in patients with thoracic surgery (41.1%, 95%CI 26.3-57.7%; I² = 74.4%) was more common than in those with gynecologic surgery (20.1%, 95%CI 15.6-25.6%; I² = 33.0%) or abdominal surgery (13.8%, 95%CI 10.3-18.4%; I² = 70.0%). Similar results were found in meta-regression. MRSA accounted for 41.3% (95%CI 36.5-46.3%; I² = 64.6%) of S. aureus, significantly lower than that in the US (P-value = 0.001). MRSA was sensitive to vancomycin (522/522) and linezolid (93/94), while 79.9% (95%CI 67.4-88.4%; I² = 0%) and 92.0% (95%CI 80.2-97.0%; I² = 0%) of MRSA was resistant to clindamycin and erythromycin respectively.

Conclusion

The overall proportion of S. aureus among SSIs in China was similar to that in the US but seemed higher than those reported through the Chinese national surveillance system. Proportions of S. aureus SSIs may vary with different surgery types. Commonly seen in SSIs, MRSA tended to be highly sensitive to vancomycin and linezolid but mostly resistant to clindamycin and erythromycin.