Regulation of Natural Killer Cell TGF-ß and AhR Signaling Pathways Via the Intestinal Microbiota is Critical for Host Defense Against Alcohol-Associated Bacterial Pneumonia.

Alcohol use is an independent risk factor for the development of bacterial pneumonia due, in part, to impaired mucus-facilitated clearance, macrophage phagocytosis, and recruitment of neutrophils. Alcohol consumption is also known to reduce peripheral natural killer (NK) cell numbers and compromises NK cell cytolytic activity, especially NK cells with a mature phenotype. However, the role of innate lymphocytes, such as NK cells during host defense against alcohol-associated bacterial pneumonia is essentially unknown. We have previously shown that indole supplementation mitigates increases in pulmonary bacterial burden and improves pulmonary NK cell recruitment in alcohol-fed mice, which were dependent of aryl hydrocarbon receptor (AhR) signaling. Employing a binge-on-chronic alcohol-feeding model we sought to define the role and interaction of indole and NK cells during pulmonary host defense against alcohol-associated pneumonia. We demonstrate that alcohol dysregulates NK cell effector function and pulmonary recruitment via alterations in two key signaling pathways. We found that alcohol increases transforming growth factor beta (TGF-ß) signaling, while suppressing AhR signaling. We further demonstrated that NK cells isolated from alcohol-fed mice have a reduced ability to kill Klebsiella pneumoniae. NK cell migratory capacity to chemokines was also significantly altered by alcohol, as NK cells isolated from alcohol-fed mice exhibited preferential migration in response to CXCR3 chemokines but exhibited reduced migration in response to CCR2, CXCR4, and CX3CR1 chemokines. Together this data suggests that alcohol disrupts NK cell specific TGF-ß and AhR signaling pathways leading to decreased pulmonary recruitment and cytolytic activity thereby increasing susceptibility to alcohol-associated bacterial pneumonia.

Human Alcohol-Microbiota Mice have Increased Susceptibility to Bacterial Pneumonia.

Preclinical studies have shown that chronic alcohol abuse leads to alterations in the gastrointestinal microbiota that are associated with behavior changes, physiological alterations, and immunological effects. However, such studies have been limited in their ability to evaluate the direct effects of alcohol-associated dysbiosis. To address this, we developed a humanized alcohol-microbiota mouse model to systematically evaluate the immunological effects of chronic alcohol abuse mediated by intestinal dysbiosis. Germ-free mice were colonized with human fecal microbiota from individuals with high and low Alcohol Use Disorders Identification Test (AUDIT) scores and bred to produce human alcohol-associated microbiota or human control-microbiota F1 progenies. F1 offspring colonized with fecal microbiota from individuals with high AUDIT scores had increased susceptibility to Klebsiella pneumoniae and Streptococcus pneumoniae pneumonia, as determined by increased mortality rates, pulmonary bacterial burden, and post-infection lung damage. These findings highlight the importance of considering both the direct effects of alcohol and alcohol-induced dysbiosis when investigating the mechanisms behind alcohol-related disorders and treatment strategies.

The Inherited Intestinal Microbiota from Myeloid-Specific ZIP8KO Mice Impairs Pulmonary Host Defense against Pneumococcal Pneumonia.

Intestinal dysbiosis increases susceptibility to infection through the alteration of metabolic profiles, which increases morbidity. Zinc (Zn) homeostasis in mammals is tightly regulated by 24 Zn transporters. ZIP8 is unique in that it is required by myeloid cells to maintain proper host defense against bacterial pneumonia. In addition, a frequently occurring ZIP8 defective variant (SLC39A8 rs13107325) is strongly associated with inflammation-based disorders and bacterial infection. In this study, we developed a novel model to study the effects of ZIP8-mediated intestinal dysbiosis on pulmonary host defense independent of the genetic effects. Cecal microbial communities from a myeloid-specific Zip8 knockout mouse model were transplanted into germ-free mice. Conventionalized ZIP8KO-microbiota mice were then bred to produce F1 and F2 generations of ZIP8KO-microbiota mice. F1 ZIP8KO-microbiota mice were also infected with S. pneumoniae, and pulmonary host defense was assessed. Strikingly, the instillation of pneumococcus into the lung of F1 ZIP8KO-microbiota mice resulted in a significant increase in weight loss, inflammation, and mortality when compared to F1 wild-type (WT)-microbiota recipients. Similar defects in pulmonary host defense were observed in both genders, although consistently greater in females. From these results, we conclude that myeloid Zn homeostasis is not only critical for myeloid function but also plays a significant role in the maintenance and control of gut microbiota composition. Further, these data demonstrate that the intestinal microbiota, independent of host genetics, play a critical role in governing host defense in the lung against infection. Finally, these data strongly support future microbiome-based interventional studies, given the high incidence of zinc deficiency and the rs13107325 allele in humans.

Black thyroid gland and tracheal cartilage.

A woman in her 70s with a history of chronic minocycline use presented with complaints of a non-tender posterior neck mass. A thyroid gland ultrasound showed a highly suspicious right thyroid nodule. A total thyroidectomy revealed darkened discolouration of the thyroid gland and tracheal cartilage. The pathology report showed dark brown granules representing melanin. Chronic minocycline usage is known to cause pigmentation of nails, teeth, bones and the thyroid gland. Our case highlights the importance of recognising that long-term use of minocycline can cause discolouration of the thyroid and tracheal cartilage. Current case studies do not show any adverse health effects associated with black thyroid and tracheal cartilage. For patients who are to undergo neck surgery, physicians need to be aware of this side effect, and that further intervention, such as surgical resection, may not be required.

A Selective and Sensitive LC-MS/MS Method for Quantitation of Indole in Mouse Serum and Tissues.

Indole is an endogenous substance currently being evaluated as a biomarker for ulcerative colitis, irritable bowel syndrome, Crohn's disease and non-alcoholic fatty liver disease. A novel, selective, and sensitive method using liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) was developed for quantitation of indole concentrations in mouse plasma and tissues. Samples were prepared by protein precipitation using ice-cold acetonitrile (ACN) followed by injecting the extracted analyte to LC-MS/MS system. Indole was separated using Synergi Fusion C18 (4 µm, 250 × 2.0 mm) column with mobile phase 0.1% aqueous formic acid (A) and methanol (B) using gradient flow with run time 12 min. The mass spectrometer was operated in atmospheric pressure chemical ionization (APCI) positive mode at unit resolution in multiple reaction monitoring (MRM) mode, using precursor ion > product ion combinations of 118.1 > 91.1 m/z for indole and 124.15 > 96.1 m/z for internal standard (IS) indole d7. The MS/MS response was linear over the range of indole concentrations (1−500 ng/mL). The validated method was applied for quantitation of indole concentrations range in mouse lungs (4.3−69.4 ng/g), serum (0.8−38.7 ng/mL) and cecum (1043.8−12,124.4 ng/g). This method would help investigate the role of indole as a biomarker and understand its implications in different disease states.

ZIP8-Mediated Intestinal Dysbiosis Impairs Pulmonary Host Defense against Bacterial Pneumonia.

Pneumococcal pneumonia is a leading cause of morbidity and mortality worldwide. An increased susceptibility is due, in part, to compromised immune function. Zinc is required for proper immune function, and an insufficient dietary intake increases the risk of pneumonia. Our group was the first to reveal that the Zn transporter, ZIP8, is required for host defense. Furthermore, the gut microbiota that is essential for lung immunity is adversely impacted by a commonly occurring defective ZIP8 allele in humans. Taken together, we hypothesized that loss of the ZIP8 function would lead to intestinal dysbiosis and impaired host defense against pneumonia. To test this, we utilized a novel myeloid-specific Zip8KO mouse model in our studies. The comparison of the cecal microbial composition of wild-type and Zip8KO mice revealed significant differences in microbial community structure. Most strikingly, upon a S. pneumoniae lung infection, mice recolonized with Zip8KO-derived microbiota exhibited an increase in weight loss, bacterial dissemination, and lung inflammation compared to mice recolonized with WT microbiota. For the first time, we reveal the critical role of myeloid-specific ZIP8 on the maintenance of the gut microbiome structure, and that loss of ZIP8 leads to intestinal dysbiosis and impaired host defense in the lung. Given the high incidence of dietary Zn deficiency and the ZIP8 variant allele in the human population, additional investigation is warranted to improve surveillance and treatment strategies.

A comparison of sagittal MRI and lateral radiography in determining the Insall-Salvati ratio and diagnosing patella alta in the pediatric knee.

BACKGROUND: The Insall-Salvati ratio is a technique for determining patellar height that relies on bony landmarks. Magnetic resonance imaging (MRI) and plain radiography are used interchangeably to assess the Insall-Salvati ratio in the pediatric population despite the lack of validity in the literature. OBJECTIVE: The purpose of this study was to investigate if the Insall-Salvati ratio and patella alta as determined on MRI are comparable to those determined on radiography in pediatric patients. MATERIALS AND METHODS: We conducted a retrospective review of 49 pediatric patients (age range: 7.5-17.0 years) with unfused growth plates who underwent both knee MRI and lateral knee radiography. Measurements for calculating the Insall-Salvati ratio (the ratio of patella tendon length to patella length) were obtained by three observers. Data were analyzed using paired t-tests and Pearson's correlation. A reliability assessment and inter-method agreements were performed. Patella alta was defined as an Insall-Salvati ratio > 1.2. Additional cutoffs of Insall-Salvati ratios > 1.3 and > 1.4 were also analyzed. RESULTS: There was no statistically significant difference between Insall-Salvati ratio as determined on MRI (mean: 1.20) and radiographs (mean: 1.25; P > 0.05). There was a strong correlation between Insall-Salvati ratio as determined on MRI and radiographs (Pearson's r = 0.6) with moderate consistency (Cronbach's alpha = 0.78). There was a good level of agreement between the diagnosis of patella alta on MRI and radiographs when defined as an Insall-Salvati ratio greater than 1.2 and 1.3 (Cohen's kappa = 0.61). CONCLUSION: The results demonstrate a strong association between Insall-Salvati ratio and patella alta derived from MRI and radiographs in children ages 7.5 years and older.

Evaluation of DNA damage and stress in wildlife chronically exposed to low-dose, low-dose rate radiation from the Fukushima Dai-ichi Nuclear Power Plant accident.

The health effects associated with chronic low-dose, low-dose rate (LD-LDR) exposures to environmental radiation are uncertain. All dose-effect studies conducted outside controlled laboratory conditions are challenged by inherent complexities of ecological systems and difficulties quantifying dose to free-ranging organisms in natural environments. Consequently, the effects of chronic LD-LDR radiation exposures on wildlife health remain poorly understood and much debated. Here, samples from wild boar (Sus scrofa leucomystax) and rat snakes (Elaphe spp.) were collected between 2016 and 2018 across a gradient of radiation exposures in Fukushima, Japan. In vivo biomarkers of DNA damage and stress were evaluated as a function of multiple measurements of radiation dose. Specifically, we assessed frequencies of dicentric chromosomes (Telomere-Centromere Fluorescence in situ Hybridization: TC-FISH), telomere length (Telo-FISH, qPCR), and cortisol hormone levels (Enzyme Immunoassay: EIA) in wild boar, and telomere length (qPCR) in snakes. These biological parameters were then correlated to robust calculations of radiation dose rate at the time of capture and plausible upper bound lifetime dose, both of which incorporated internal and external dose. No significant relationships were observed between dicentric chromosome frequencies or telomere length and dose rate at capture or lifetime dose (p value range: 0.20-0.97). Radiation exposure significantly associated only with cortisol, where lower concentrations were associated with higher dose rates (r2 = 0.58; p < 0.0001), a relationship that was likely due to other (unmeasured) factors. Our results suggest that wild boar and snakes chronically exposed to LD-LDR radiation sufficient to prohibit human occupancy were not experiencing significant adverse health effects as assessed by biomarkers of DNA damage and stress.

Erratum: Author Correction: LepVax, a defined subunit vaccine that provides effective pre-exposure and post-exposure prophylaxis of M. leprae infection.

[This corrects the article DOI: 10.1038/s41541-018-0050-z.].

LepVax, a defined subunit vaccine that provides effective pre-exposure and post-exposure prophylaxis of M. leprae infection.

Sustained elimination of leprosy as a global health concern likely requires a vaccine. The current standard, BCG, confers only partial protection and precipitates paucibacillary (PB) disease in some instances. When injected into mice with the T helper 1 (Th1)-biasing adjuvant formulation Glucopyranosyl Lipid Adjuvant in stable emulsion (GLA-SE), a cocktail of three prioritized antigens (ML2055, ML2380 and ML2028) reduced M. leprae infection levels. Recognition and protective efficacy of a single chimeric fusion protein incorporating these antigens, LEP-F1, was confirmed in similar experiments. The impact of post-exposure immunization was then assessed in nine-banded armadillos that demonstrate a functional recapitulation of leprosy. Armadillos were infected with M. leprae 1 month before the initiation of post-exposure prophylaxis. While BCG precipitated motor nerve conduction abnormalities more rapidly and severely than observed for control infected armadillos, motor nerve injury in armadillos treated three times, at monthly intervals with LepVax was appreciably delayed. Biopsy of cutaneous nerves indicated that epidermal nerve fiber density was not significantly altered in M. leprae-infected animals although Remak Schwann cells of the cutaneous nerves in the distal leg were denser in the infected armadillos. Importantly, LepVax immunization did not exacerbate cutaneous nerve involvement due to M. leprae infection, indicating its safe use. There was no intraneural inflammation but a reduction of intra axonal edema suggested that LepVax treatment might restore some early sensory axonal function. These data indicate that post-exposure prophylaxis with LepVax not only appears safe but, unlike BCG, alleviates and delays the neurologic disruptions caused by M. leprae infection.

Intraoperative optical coherence tomography of the human thyroid: Feasibility for surgical assessment.

Thyroid nodules assessed with ultrasound and fine-needle aspiration biopsy are diagnosed as "suspicious" or "indeterminate" in 15%-20% of the cases. Typically, total thyroidectomy is performed in such cases; however, only 25%-50% are found to be cancerous upon final histopathologic analysis. Here we demonstrate optical coherence tomography (OCT) imaging of the human thyroid as a potential intraoperative imaging tool for providing tissue assessment in real time during surgical procedures. Fresh excised tissue specimens from 28 patients undergoing thyroid surgery were imaged in the laboratory using a benchtop OCT system. Three-dimensional OCT images showed different microstructural features in normal, benign, and malignant thyroid tissues. A similar portable OCT system was then designed and constructed for use in the operating room, and intraoperative imaging of excised thyroid tissue from 6 patients was performed during the surgical procedure. The results demonstrate the potential of OCT to provide real-time imaging guidance during thyroid surgeries.

Effects of Human ES-Derived Neural Stem Cell Transplantation and Kindling in a Rat Model of Traumatic Brain Injury.

Traumatic brain injury (TBI) is one of the leading causes of death and disability in the population worldwide, with a broad spectrum of symptoms and disabilities. Posttraumatic hyperexcitability is one of the most common neurological disorders that affect people after a head injury. A reliable animal model of posttraumatic hyperexcitability induced by TBI which allows one to test effective treatment strategies is yet to be developed. To address these issues, in the present study, we tested human embryonic stem cell-derived neural stem cell (NSC) transplantation in an animal model of posttraumatic hyperexcitability in which the brain injury was produced in one hemisphere of immunodeficient athymic nude rats by controlled cortical impact, and spontaneous seizures were produced by repeated electrical stimulation (kindling) in the contralateral hemisphere. At 14 wk posttransplantation, we report human NSC (hNSC) survival and differentiation into all 3 neural lineages in both sham and injured animals. We observed twice as many surviving hNSCs in the injured versus sham brain, and worse survival on the kindled side in both groups, indicating that kindling/seizures are detrimental to survival or proliferation of hNSCs. We also replicated our previous finding that hNSCs can ameliorate deficits on the novel place recognition task,33 but such improvements are abolished following kindling. We found no significant differences pre- or post-kindling on the elevated plus maze. No significant correlations were observed between hNSC survival and cognitive performance on either task. Together these findings suggest that Shef6-derived hNSCs may be beneficial as a therapy for TBI, but not in animals or patients with posttraumatic hyperexcitability.

Cutaneous nerve biomarkers in transthyretin familial amyloid polyneuropathy.

OBJECTIVE: To determine the utility of skin biopsies as a biomarker of disease severity in subjects with amyloid neuropathy. METHODS: Five groups of patients were studied: (1) transthyretin (TTR) familial amyloidotic polyneuropathy (FAP; n = 20), (2) TTR mutation carriers without peripheral neuropathy (TTR-noPN; n = 10), (3) healthy controls (n = 20), (4) diabetic neuropathy disease controls (n = 20), and (5) patients with light-chain (AL) amyloid (n = 2). All subjects underwent neurological examination and 3mm skin biopsies. Sections were stained with anti-PGP9.5, anti-TTR, and Congo red. Intraepidermal (IENFD), sweat gland (SGNFD), and pilomotor nerve fiber densities (PMNFD) were measured. Correlations between the amount of amyloid present (amyloid burden), fiber subtype, and Neuropathy Impairment Score in the Lower Limbs (NIS-LL) were evaluated. RESULTS: IENFD, SGNFD, and PMNFD were all significantly reduced in TTR-FAP patients versus healthy controls, whereas TTR-noPN subjects had intermediate reductions. Lower nerve fiber densities were associated with NIS-LL (p < 0.001). Congo red staining revealed brilliant red amyloid deposits confirmed by apple-green birefringence within dermal collagen, sweat glands, and arrector pili that engulfed axons. The diagnostic sensitivity and specificity to detect amyloid in skin were 70% and 100%. Both AL amyloidosis and 2 of 10 TTR-noPN subjects were Congo red-positive. Amyloid burden correlated with IENFD (r = -0.63), SGNFD (r = -0.67), PMNFD (r = -0.50), and NIS-LL (r = -0.57). Wild-type TTR staining was less prominent in TTR-FAP patients. INTERPRETATION: Cutaneous amyloid was detected in 70% of TTR-FAP and 20% of TTR-noPN subjects. Amyloid burden correlated strongly with reductions in IENFD, SGNFD, PMNFD, and NIS-LL. Skin is an attractive tissue to establish an amyloid diagnosis, and amyloid burden has potential as a biomarker to detect treatment effect in TTR-FAP drug trials. Ann Neurol 2017;82:44-56.

Benign sacrococcygeal teratoma incidentally found on routine scoliosis radiographs in a 12-year-old female: a case report.

OBJECTIVE: To describe the imaging characteristics of sacrococcygeal teratomas and to review appropriate diagnostic evaluation and management. CLINICAL FEATURES: 12-year-old otherwise healthy, asymptomatic female with an incidental pelvic mass found on routine scoliosis radiographs. INTERVENTION AND OUTCOME: The pelvic mass was further evaluated by MRI and CT scan. Management consisted of successful surgical resection with no postoperative complications. Pathology confirmed a mature, benign, sacrococcygeal teratoma. CONCLUSIONS: Chiropractors manage patients with scoliosis, which may include radiographic surveillance. Familiarity with the radiographic features of masses such as sacrococcygeal teratomas is important for prompt diagnosis and management.

OBJECTIF: Décrire les caractéristiques d'imagerie des tératomes sacrococcygiens et examiner l'évaluation et la prise en charge du diagnostic approprié. CARACTÉRISTIQUES CLINIQUES: Jeune fille asymptomatique de 12 ans en bonne santé avec une masse pelvienne découverte fortuitement après des radiographies de routine pour la scoliose. INTERVENTION ET RÉSULTATS: La masse pelvienne a davantage été évaluée à partir d'images d'IRM et de TDM. La solution était la résection chirurgicale réussie sans complications postopératoires. La pathologie a confirmé un tératome sacrococcygien mature bénin. CONCLUSIONS: Les chiropraticiens prennent en charge les patients atteints de scoliose, ce qui peut inclure la surveillance radiographique. La familiarité avec les caractéristiques radiographiques des masses, comme les tératomes sacrococcygiens, est importante pour leur diagnostic et la prise en charge rapide.

Transplantation of human neural stem cells restores cognition in an immunodeficient rodent model of traumatic brain injury.

Traumatic brain injury (TBI) in humans can result in permanent tissue damage and has been linked to cognitive impairment that lasts years beyond the initial insult. Clinically effective treatment strategies have yet to be developed. Transplantation of human neural stem cells (hNSCs) has the potential to restore cognition lost due to injury, however, the vast majority of rodent TBI/hNSC studies to date have evaluated cognition only at early time points, typically <1month post-injury and cell transplantation. Additionally, human cell engraftment and long-term survival in rodent models of TBI has been difficult to achieve due to host immunorejection of the transplanted human cells, which confounds conclusions pertaining to transplant-mediated behavioral improvement. To overcome these shortfalls, we have developed a novel TBI xenotransplantation model that utilizes immunodeficient athymic nude (ATN) rats as the host recipient for the post-TBI transplantation of human embryonic stem cell (hESC) derived NSCs and have evaluated cognition in these animals at long-term (≥2months) time points post-injury. We report that immunodeficient ATN rats demonstrate hippocampal-dependent spatial memory deficits (Novel Place, Morris Water Maze), but not non-spatial (Novel Object) or emotional/anxiety-related (Elevated Plus Maze, Conditioned Taste Aversion) deficits, at 2-3months post-TBI, confirming that ATN rats recapitulate some of the cognitive deficits found in immunosufficient animal strains. Approximately 9-25% of transplanted hNSCs survived for at least 5months post-transplantation and differentiated into mature neurons (NeuN, 18-38%), astrocytes (GFAP, 13-16%), and oligodendrocytes (Olig2, 11-13%). Furthermore, while this model of TBI (cortical impact) targets primarily cortex and the underlying hippocampus and generates a large lesion cavity, hNSC transplantation facilitated cognitive recovery without affecting either lesion volume or total spared cortical or hippocampal tissue volume. Instead, we have found an overall increase in host hippocampal neuron survival in hNSC transplanted animals and demonstrate that a correlation exists between hippocampal neuron survival and cognitive performance. Together, these findings support the use of immunodeficient rodents in models of TBI that involve the transplantation of human cells, and suggest that hNSC transplantation may be a viable, long-term therapy to restore cognition after brain injury.

A Kinect-based system for automatic recording of some pigeon behaviors.

Contact switches and touch screens are the state of the art for recording pigeons' pecking behavior. Recording other behavior, however, requires a different sensor for each behavior, and some behaviors cannot easily be recorded. We present a flexible and inexpensive image-based approach to detecting and counting pigeon behaviors that is based on the Kinect sensor from Microsoft. Although the system is as easy to set up and use as the standard approaches, it is more flexible because it can record behaviors in addition to key pecking. In this article, we show how both the fast, fine motion of key pecking and the gross body activity of feeding can be measured. Five pigeons were trained to peck at a lighted contact switch, a pigeon key, to obtain food reward. The timing of the pecks and the food reward signals were recorded in a log file using standard equipment. The Kinect-based system, called BehaviorWatch, also measured the pecking and feeding behavior and generated a different log file. For key pecking, BehaviorWatch had an average sensitivity of 95% and a precision of 91%, which were very similar to the pecking measurements from the standard equipment. For detecting feeding activity, BehaviorWatch had a sensitivity of 95% and a precision of 97%. These results allow us to demonstrate that an advantage of the Kinect-based approach is that it can also be reliably used to measure activity other than key pecking.

The importance of empiric abdominal computed tomography after urgent laparotomy for trauma: do they reveal unexpected injuries?

BACKGROUND: Many institutions now use empiric full-body computed tomography (CT) as a standard step in the initial workup of stable trauma patients. Recent data suggest that these scans may reveal unexpected injuries and improve survival in patients with polytrauma. However, patients who are unstable on presentation are often taken to the operating room (OR) without CT. Many of these patients undergo empiric full-body CTs after being stabilized in the OR, yet few data exist regarding how often early postoperative CT reveals unexpected injuries within compartments that have been explored surgically. Thus, the objective of this study was to determine if empiric abdominal/pelvic (ABD) CT after emergent trauma laparotomies are likely to reveal missed injuries requiring urgent management and improve patient management compared with clinical judgment alone. METHODS: We review retrospectively 496 trauma patients who required urgent exploratory laparotomy at UPMC Presbyterian Hospital from 2007 to 2011. Patients were included if they went to the OR for exploratory laparotomy directly from the emergency department within 2 hours of arrival. Patients were excluded if they received any preoperative ABD CT imaging. Patients who expired in the OR were similarly excluded. Patients were stratified into 2 groups based on whether or not they received an empiric ABD CT in the 24 hours immediately after laparotomy. Medical records were reviewed to look for differences in missed injuries, urgent reexplorations, nontherapeutic interventions, and time to urgent reexploration. RESULTS: There were 278 patients who met inclusion at exclusion criteria and constituted the study cohort. Of these patients, 124 underwent early empiric postoperative ABD CT imaging (45%). The remaining 154 patients did not undergo early ABD imaging (no CT group). The overall cohort had a 45% incidence of damage control procedures and a 9% rate of negative laparotomy. The 2 groups were statistically similar in age, presenting vitals, and abdominal Abbreviated Injury Scores. When the ABD CT group was compared with the no CT group, there was no difference in the overall rate of urgent reexplorations (7.3 vs 7.1%; P = .956), nontherapeutic urgent reexplorations (22 vs 18%; P = .822), or time to urgent reexploration (14 ± 10 vs 12 ± 10 hours; P = .686). Out of the 124 ABD CT patients, only 5 (4.0%) were diagnosed with injuries that were not identified at the time of the initial operation or caused by operative technique. When controlling for demographics, mechanism of injury, and injury severity, a logistic regression analysis revealed that early postoperative ABD CT was not associated with any differential risk of the need for further intervention (odds ratio, 0.85; 95% CI, 0.37-1.9; P = .691). CONCLUSION: The use of ABD CT soon after trauma laparotomy did not provide meaningful improvements in patient care in the cohort studied. Further higher level research is needed to clarify what role empiric ABD CT should play in the early postoperative period.

Basic flap design.

Basic flap design utilization for reconstruction of head and neck defects requires creativity from the surgeon. Ultimately, the surgeon must closely restore the basic functions and properties of the surgical flap and adjacent tissue. All options within the reconstructive ladder should be considered. When possible, like should be replaced with like (similar tissue) within an esthetic zone. When considering a flap design, the surgeon must remember that the donor site must be closed in an esthetic and functional manner. Finally, knowledge of normal anatomy, the extent of the defect, and the patient is vital for successful outcomes.

Progesterone activates multiple innate immune pathways in Chlamydia trachomatis-infected endocervical cells.

PROBLEM: Susceptibility to Chlamydia trachomatis infection is increased by oral contraceptives and modulated by sex hormones. We therefore sought to determine the effects of female sex hormones on the innate immune response to C. trachomatis infection. METHOD OF STUDY: ECC-1 endometrial cells, pre-treated with oestradiol or progesterone, were infected with C. trachomatis and the host transcriptome analysed by Illumina Sentrix HumanRef-8 microarray. Primary endocervical epithelial cells, prepared at either the proliferative or secretory phase of the menstrual cycle, were infected with C. trachomatis and cytokine gene expression determined by quantitative RT-PCR analysis. RESULTS: Chlamydia trachomatis yield from progesterone-primed ECC-1 cells was significantly reduced compared with oestradiol-treated cells. Genes upregulated in progesterone-treated and Chlamydia-infected cells only included multiple CC and CXC chemokines, IL-17C, IL-29, IL-32, TNF-α, DEFB4B, LCN2, S100A7-9, ITGAM, NOD2, JAK1, IL-6ST, type I and II interferon receptors, numerous interferon-stimulated genes and STAT6. CXCL10, CXCL11, CX3 CL1 and IL-17C, which were also upregulated in infected secretory-stage primary cells, and there was a trend towards higher levels of immune mediators in infected secretory-phase compared with proliferative-phase cells. CONCLUSION: Progesterone treatment primes multiple innate immune pathways in hormone-responsive epithelial cells that could potentially increase resistance to chlamydial infection.

The IL-6 response to Chlamydia from primary reproductive epithelial cells is highly variable and may be involved in differential susceptibility to the immunopathological consequences of chlamydial infection.

BACKGROUND: Chlamydia trachomatis infection results in reproductive damage in some women. The process and factors involved in this immunopathology are not well understood. This study aimed to investigate the role of primary human cellular responses to chlamydial stress response proteases and chlamydial infection to further identify the immune processes involved in serious disease sequelae. RESULTS: Laboratory cell cultures and primary human reproductive epithelial cultures produced IL-6 in response to chlamydial stress response proteases (CtHtrA and CtTsp), UV inactivated Chlamydia, and live Chlamydia. The magnitude of the IL-6 response varied considerably (up to 1000 pg ml(-1)) across different primary human reproductive cultures. Thus different levels of IL-6 production by reproductive epithelia may be a determinant in disease outcome. Interestingly, co-culture models with either THP-1 cells or autologous primary human PBMC generally resulted in increased levels of IL-6, except in the case of live Chlamydia where the level of IL-6 was decreased compared to the epithelial cell culture only, suggesting this pathway may be able to be modulated by live Chlamydia. PBMC responses to the stress response proteases (CtTsp and CtHtrA) did not significantly vary for the different participant cohorts. Therefore, these proteases may possess conserved innate PAMPs. MAP kinases appeared to be involved in this IL-6 induction from human cells. Finally, we also demonstrated that IL-6 was induced by these proteins and Chlamydia from mouse primary reproductive cell cultures (BALB/C mice) and mouse laboratory cell models. CONCLUSIONS: We have demonstrated that IL-6 may be a key factor for the chlamydial disease outcome in humans, given that primary human reproductive epithelial cell culture showed considerable variation in IL-6 response to Chlamydia or chlamydial proteins, and that the presence of live Chlamydia (but not UV killed) during co-culture resulted in a reduced IL-6 response suggesting this response may be moderated by the presence of the organism.