Bladder Pain Sensitivity Is a Potential Risk Factor for Irritable Bowel Syndrome.

BACKGROUND: Although dysmenorrhea is a highly prevalent risk factor for irritable bowel syndrome (IBS), the factors underlying this risk are not fully understood. Prior studies support a hypothesis that repeated distressing menstrual pain promotes cross-organ pelvic sensitization with heightened visceral sensitivity. AIMS: To further explore cross-organ pelvic sensitization we examined the association of dysmenorrhea, provoked bladder pain, and other putative factors with self-reported IBS-domain pain frequency and new onset after 1-year follow up. METHODS: We measured visceral pain sensitivity with a noninvasive provoked bladder pain test in a cohort of reproductive-aged women, enriched for those reporting moderate-to-severe menstrual pain intensity but without any prior IBS diagnosis (n = 190). We analyzed the relationship between menstrual pain, provoked bladder pain, pain catastrophizing, anxiety, and depression with primary outcomes: (1) frequency of self-reported IBS-domain pain and (2) new onset of IBS-domain pain after 1-year follow up. RESULTS: All hypothesized factors correlated with the frequency of IBS-domain pain (p's ≤ 0.038). In a cross-sectional model, only menstrual pain (standardized adjusted odds ratio 2.07), provoked bladder pain (1.49), and anxiety (1.90) were independently associated with IBS-domain pain ≥ 2 days/month (C statistic = 0.79). One year later, provoked bladder pain (3.12) was the only significant predictor of new onset IBS-domain pain (C statistic = 0.87). CONCLUSION: Increased visceral sensitivity among women with dysmenorrhea could lead to IBS. Because provoked bladder pain predicted subsequent IBS, prospective studies should be performed to see if the early treatment of visceral hypersensitivity mitigates IBS.

Potential mechanisms contributing to the high cadmium removal efficiency from contaminated soil by using effective microorganisms as novel electrolyte in electrokinetic remediation applications.

In this study, we tested the ability of a solution of effective microorganisms (EM) to remove cadmium from soil. Experimental results revealed that EM had an overall cadmium removal efficiency of 90.5% after 7 days of electrokinetic (EK) treatment. During EK treatment, EM exhibited a low initial pH of 3.6 and a high conductivity of 7.0 mS/m; therefore, they reduced the pH of the anode after an electric field was applied. EM had a surface tension of 50.3 dyne/cm and exhibited biosurfactant property in the EK experiments. The cadmium removal efficiency of EM in soil was compared with that of tap water, citric acid, and ethylenediaminetetraacetic acid (EDTA). The results revealed that after 7 days of EK treatment, EM had a higher cadmium removal efficiency than did citric acid (72.3%), EDTA (75.4%), and tap water (21.7%). This result can be partly attributed to the biosurfactant property of EM, which enables them to penetrate deeply into the soil matrix and thus dissolve a high quantity of pollutants. Overall, the results of this study indicate that EM can serve as an economic and efficient biosurfactant for removing cadmium from soil in EK applications.

Three-month Chan-Chuang qigong program improves physical performance and quality of life of patients with cognitive impairment: A randomized controlled trial.

This randomized controlled trial was conducted to evaluate the effects of a 3-month-long Chan-Chuang qigong program on patients' physical performance and quality of life while excluding the influence caused by the progression of their cognitive impairment. Patients with mild to moderate cognitive impairment were recruited from two dementia daycare centers in Taiwan. The control group (n = 41) received the standardized plan of treatment, and the qigong group (n = 39) received the standardized plan of treatment plus the Chan-Chuang qigong program. The outcomes were muscle strength, muscle endurance, exercise capacity, and quality of life. After controlling for the progression of cognitive impairment, the qigong group showed significant improvements over the control group and baseline in muscle strength and exercise capacity at Months 2 and 3 (p < 0.05) and in muscle endurance at Months 1, 2, and 3 (p < 0.05). The Cognitron test scores were significantly associated with muscle strength (p = 0.03), whereas the Corsi block-tapping test scores were significantly associated with exercise capacity (p = 0.001). Furthermore, a significant between-group difference was detected in the physical (p = 0.01), not mental (p = 0.83), component of quality of life. The 3-month Chan-Chuang qigong program can be applied for patients with mild to moderate cognitive impairment as complementary therapy to improve their muscle strength, muscle endurance, exercise capacity, and physical quality of life. This program should be practiced for at least 2 months to achieve satisfactory results.

Clinical outcomes in ovarian cancer patients receiving three versus more cycles of chemotherapy after neoadjuvant treatment and interval cytoreductive surgery.

OBJECTIVES: To compare clinical outcomes for stage IIIC and IV ovarian cancer patients receiving neoadjuvant chemotherapy and interval cytoreductive surgery followed by up to three versus more cycles of post-operative chemotherapy. METHODS: We conducted a multi-institution retrospective cohort study of patients treated from January 2005 to February 2016 with neoadjuvant platinum-based therapy followed by interval surgery and post-operative chemotherapy. The following were exclusion criteria: more than four cycles of neoadjuvant chemotherapy, bevacizumab with neoadjuvant chemotherapy, non-platinum therapy, prior chemotherapy, and elevated CA125 values after three post-operative chemotherapy cycles. Progression-free and overall survival and toxicity profiles were compared between groups receiving up to three cycles versus more that three cycles post-operatively. RESULTS: A total of 100 patients met inclusion criteria: 41 received up to three cycles and 59 received more than three cycles. The groups were similar in terms of age, body mass index, performance status, tumor histology, optimal cytoreduction rates, and median number of neoadjuvant chemotherapy cycles. Median progression-free survival was 14 vs 16.6 months in those receiving up to three cycles versus more than three cycles, respectively (HR 0.99, 95% CI 0.58 to 1.68, p=0.97). Similarly, median overall survival was not different at 47.1 vs 69.4 months, respectively (HR 1.96, 95% CI 0.87 to 4.42, p=0.10). There were no differences in grade 2 or higher chemotherapy-related toxicities. CONCLUSIONS: Extending post-operative chemotherapy beyond three cycles in patients receiving neoadjuvant chemotherapy and interval cytoreductive surgery with normalization of CA125 levels was not associated with improved survival or greater toxicity. Future study in a larger cohort is warranted to define optimal length of cytotoxic treatment.

The effect of contralateral prophylactic mastectomy on breast-related charges: A 5-year analysis.

BACKGROUND AND OBJECTIVES: The purpose of this study was to determine charges following unilateral mastectomy (UM) and bilateral mastectomy (BM) for patients with unilateral breast cancer (UBC). We hypothesized that BM may be associated with fewer charges over time. METHODS: A retrospective review was conducted of patients with UBC treated between 2006 and 2010 with UM and BM in a large healthcare system. Institutional billing data were investigated for 5 years postoperatively to calculate the immediate and subsequent charges of all inpatient and outpatient breast-related care associated with the initial diagnosis for a subset of patients identified using propensity score matching method. RESULTS: A subset of matched patients (n = 320) undergoing UM (n = 160) or BM (n = 160) were included in this analysis. At 1 year, there was a trend toward lower total charges following UM as compared with BM (median, $125 230 vs $138 467; P = .6075). However, during years 2 to 5, total charges were significantly higher following UM vs BM ($22 128 vs $13 478; P = .0116). CONCLUSIONS: While initially higher, overall charges for BM are lower than UM between 2 and 5 years out from surgery. Further study is necessary to determine if this trend is sustained over the long term. These data can inform patient decision making regarding mastectomy for their breast cancer.

X-ray reflectivity and grazing incidence diffraction studies of interaction between human adhesion/growth-regulatory galectin-1 and DPPE-GM1 lipid monolayer at an air/water interface.

The specific interaction of ganglioside GM1 with the homodimeric (prototype) endogenous lectin galectin-1 triggers growth regulation in tumor and activated effector T cells. This proven biorelevance directed interest to studying association of the lectin to a model surface, i.e. a 1,2-dihexadecanoyl-sn-glycero-3-phosphoethanolamine/ganglioside GM1 (80 : 20 mol%) monolayer, at a bioeffective concentration. Surface expansion by the lectin insertion was detected at a surface pressure of 20 mN/m. On combining the methods of grazing incidence X-ray diffraction and X-ray reflectivity, a transient decrease in lipid-ordered phase of the monolayer was observed. The measured electron density distribution indicated that galectin-1 is oriented with its long axis in the surface plane, ideal for cis-crosslinking. The data reveal a conspicuous difference to the way the pentameric lectin part of the cholera toxin, another GM1-specific lectin, is bound to the monolayer. They also encourage further efforts to monitor effects of structurally different members of the galectin family such as the functionally antagonistic chimera-type galectin-3.

How often are interfacility transfers of spine injury patients truly necessary?

BACKGROUND CONTEXT: Traumatic spine injuries are often transferred to regional tertiary trauma centers from outside hospitals (OSHs) and subsequently discharged from the trauma center's emergency department (ED) suggesting secondary overtriage of such injuries. PURPOSE: The aim of the study was to investigate the definitive treatment and disposition of traumatic spine injuries transferred from OSH, particularly those without other trauma injuries or neurologic symptoms. STUDY DESIGN: This was a retrospective study. PATIENT SAMPLE: Adult patients presenting to a single Level 1 trauma center with spine injuries were included. OUTCOME MEASURES: The outcome measures considered in the study were appropriateness of transfer, treatment, and cost. METHODS: Four thousand five-hundred consecutive adult patients presenting to a single Level 1 trauma center with spine injuries (isolated or polytrauma) were reviewed. This consisted of 1,427 patients (32%) transferred from an OSH ED. All OSH, emergency medical services, and receiving institution (RI) patient records and imaging were reviewed. RESULTS: Patients who were neurologically intact, nonpolytrauma, and without critical medical issues at the OSH (isolated intact spine transfers) comprised 29% of transfers. Helicopters transported 13% of these patients. The most frequent injuries were compression (26%), burst (17%), and transverse process (10%) fractures. Seventy-eight percent were discharged directly from the RI's ED. Similarly, 15% were not given any formal treatment, 13% had surgery, and 72% given orthosis treatment. The average cost for transportation and ED costs for those discharged from the RI ED were $1,863 and $12,895, respectively. Of the isolated intact spine transfers, 42% were considered to be inappropriate to warrant transfer. This was defined as those sent from an OSH with an orthopedic or neurosurgeon on staff and clearly stable injuries with minimal chance of progressing to instability. Isolated intact spine transfers whose OSH spine imaging was not considered unstable was 25% of transfers with a helicopter used to transport 14% of these patients. Eighty-seven percent were discharged from the ED, whereas only 3% went onto surgery. CONCLUSIONS: This study is the first to investigate interfacility transfers with spine injuries and found high rate of secondary overtriage of neurologically intact patients with isolated spine injuries. Potential solutions include increasing spine coverage in community EDs, increasing direct communication between the OSH and the spine specialist at the tertiary center, and utilization of teleradiology.

Repeat spine imaging in transferred emergency department patients.

STUDY DESIGN: Retrospective study. OBJECTIVE: Assess frequency of repeat spine imaging in patients transferred with known spine injuries from outside hospital (OSH) to tertiary receiving institution (RI). SUMMARY OF BACKGROUND DATA: Unnecessary repeat imaging after transfer has started to become a recognized problem with the obvious issues related to repeat imaging along with potential for iatrogenic injury with movement of patients with spine problems. METHODS: Consecutive adult patients presenting to a single 1-level trauma center with spine injuries during a 51-month period were reviewed (n = 4500), resulting in 1427 patients transferred from OSH emergency department. All imaging and radiology reports from the OSH were reviewed, as well as studies performed at RI. A repeat was the same imaging modality used on the same spine region as OSH imaging. RESULTS: The overall rate of repeat spine imaging for both OSH imaging sent and not sent was 23%, and 6% if repeat spine imaging via traumagram (partial/full-body computed tomography [CT]) was excluded as a repeat. The overall rate of repeat CT was 29% (7% dedicated spine CT scans and 22% part of nondedicated spine CT scan).An observation of only those patients with OSH imaging that was sent and viewable revealed that 23% underwent repeat spine imaging with 23% undergoing repeat spine CT and 41% repeat magnetic resonance imaging.In those patients with sent and viewable OSH imaging, a lack of reconstructions prompted 14% of repeats, whereas inadequate visualization of injury site prompted 8%. In only 8% of the repeats did it change management or provide necessary surgical information. CONCLUSION: This study is the first to investigate the frequency of repeat spine imaging in transfers with known spine injuries and found a substantially high rate of repeat spine CT with minimal alteration in care. Potential solutions include only performing scans at the OSH necessary to establish a diagnosis requiring transfer and improving communication between OSH and RI physicians. LEVEL OF EVIDENCE: 4.

Portal vein infusion chemotherapy with gemcitabine after surgery for pancreatic cancer.

PURPOSES: Pancreatic cancer still has a poor prognosis even after curative resection because of the high incidence of postoperative liver metastasis. This study prospectively evaluated the feasibility and tolerability of portal vein infusion chemotherapy of gemcitabine (PVIG) as an adjuvant setting after pancreatic resection. METHODS: Thirteen patients enrolled in this study received postoperative chemotherapy with PVIG. The patients received intermittent administration of gemcitabine (800 mg/m(2)) via the portal vein on days 1, 8, and 15 after surgery. The tolerability and the toxicity of PVIG were closely monitored. RESULTS: The PVIG was started on an average of 3.1 days after surgery. Complete doses of chemotherapy (three sessions of portal infusion) were accomplished in 11 of the 13 patients. Grade 3 or 4 leukocytopenia was observed in three patients (23 %), and liver dysfunction was found in one patient (7.7 %). Grade 2 sepsis developed in two cases due to bloodstream infection. Liver metastasis was the first site of recurrence in only two patients. CONCLUSIONS: PVIG can be administered to the liver with acceptable toxicity, but myelosuppression is similar to the systemic use of gemcitabine. Careful observation is required even for locoregional chemotherapy.

Current evidence of acupuncture on polycystic ovarian syndrome.

OBJECTIVE: This paper aims to provide a literature review on evaluating the efficacy of acupuncture therapy in the treatment of polycystic ovarian syndrome (PCOS) by reviewing clinical trials; randomised and non-randomised and observational studies on PCOS. The paper will also determine the possible mechanism of acupuncture treatment in PCOS, limitations of recruited studies and suggest further improvements in future studies. DESIGN: A comprehensive literature search was conducted through the databases Medline, PubMed, EMBASE, Cochrane, AMED (Allied and Complementary Medicine), NCCAM (The National Centre for Complementary and Alternative Medicine) to identify relevant monographs. RESULTS: Four studies were recruited. Several studies showed that acupuncture significantly increases beta-endorphin levels for periods up to 24 h and may have regulatory effect on FSH, LH and androgen. beta-endorphin increased levels secondary to acupuncture affects the hyperthalamic-pituitary-adrenal (HPA) axis through promoting the release of ACTH through stimulation of its precursor pro-opiomelanocortin synthesis. INCLUSION CRITERIA: All available acupuncture studies on human subjects with PCOS from June 1970 to June 2009. EXCLUSION CRITERIA: Studies not meeting the inclusion criteria, published in languages other than English or animal studies. CONCLUSION: Acupuncture is a safe and effective treatment to PCOS as the adverse effects of pharmacologic interventions are not expected by women with PCOS. Acupuncture therapy may have a role in PCOS by: increasing of blood flow to the ovaries, reducing of ovarian volume and the number of ovarian cysts, controlling hyperglycaemia through increasing insulin sensitivity and decreasing blood glucose and insulin levels, reducing cortisol levels and assisting in weight loss and anorexia. However, well-designed, randomised controlled trials are needed to elucidate the true effect of acupuncture on PCOS.

The impact of individual human immunodeficiency virus type 1 protease mutations on drug susceptibility is highly influenced by complex interactions with the background protease sequence.

The requirement for multiple mutations for protease inhibitor (PI) resistance necessitates a better understanding of the molecular basis of resistance development. The novel bioinformatics resistance determination approach presented here elaborates on genetic profiles observed in clinical human immunodeficiency virus type 1 (HIV-1) isolates. Synthetic protease sequences were cloned in a wild-type HIV-1 background to generate a large number of close variants, covering 69 mutation clusters between multi-PI-resistant viruses and their corresponding genetically closely related, but PI-susceptible, counterparts. The vast number of mutants generated facilitates a profound and broad analysis of the influence of the background on the effect of individual PI resistance-associated mutations (PI-RAMs) on PI susceptibility. Within a set of viruses, all PI-RAMs that differed between susceptible and resistant viruses were varied while maintaining the background sequence from the resistant virus. The PI darunavir was used to evaluate PI susceptibility. Single sets allowed delineation of the impact of individual mutations on PI susceptibility, as well as the influence of PI-RAMs on one another. Comparing across sets, it could be inferred how the background influenced the interaction between two mutations, in some cases even changing antagonistic relationships into synergistic ones or vice versa. The approach elaborates on patient data and demonstrates how the specific mutational background greatly influences the impact of individual mutations on PI susceptibility in clinical patterns.

Long-term survival after hepatectomy for hepatic recurrence of carcinoma of the papilla of Vater.

We present herein a rare case of a long-term survivor after major hepatectomy performed for a metastatic liver tumor from carcinoma of the papilla of Vater. A 74-year-old man had undergone a pancreaticoduodenectomy for carcinoma of the papilla of Vater with obstructive jaundice, in April 1995. Histologically, an exposed mass-forming type of tumor, measuring 40 x 30 mm, was composed of mucinous and papillary adenocarcinoma, invading into the muscularis propria of the duodenum, without lymph node metastases. The patient did not receive any type of chemotherapy. In September 1999, a solitary hepatic tumor, 3.5 cm in diameter, was detected in segment VIII of the liver by computed tomography. In November 1999, right hepatic lobectomy was carried out. The anterior and posterior portal pedicles were ligated and dissected in the hepatic parenchyma so as not to compromise the hepaticojejunostomy. After the hepatectomy, the patient was treated with low-dose tegafur/uracil/cisplatin therapy for approximately 2 years. He has been doing well, without recurrence, for 5 years after the hepatectomy. Hepatectomy for hepatic metastases from carcinoma of the papilla of Vater is thought to be a useful surgical treatment in selected patients.

Increase in laminin expression in allergic airway remodelling and decrease by dexamethasone.

Lung expression of the extracellular matrix protein, laminin, and its receptor, laminin-1 receptor, were examined in a mouse model of asthma with airway remodelling. Ovalbumin (OVA) was administered to BALB/c mice, intraperitoneally on days 0 and 14, and intranasally periodically between days 14 and 75. The mice developed airway eosinophil and mononuclear inflammatory cell infiltration and fibrosis. On day 76, a marked increase in total laminin was seen in the airways of OVA-treated mice compared to controls by Western blot analysis. The increased laminin expression was detected immunocytochemically in the thickened subepithelial basement membrane and around airways and blood vessels. The OVA-treated mice showed increased expression of the alpha1, beta1, and gamma1 chains of the laminin-1 isoform in monocytes, macrophages and eosinophils infiltrating the airways. Laminin-1 receptor expression was increased in inflammatory and endothelial cells in the lungs of OVA-treated mice compared to controls. Treatment of OVA-sensitised/challenged mice with dexamethasone reduced airway expression of laminin and laminin-1 receptor in OVA-treated mice but not airway hyperresponsiveness to methacholine. Laminin deposition may be an important component of the airway remodelling observed in chronic allergic lung inflammation and is a process modulated by corticosteroids.

Impact of topical oils on the skin barrier: possible implications for neonatal health in developing countries.

UNLABELLED: Topical therapy to enhance skin barrier function may be a simple, low-cost, effective strategy to improve outcome of preterm infants with a developmentally compromised epidermal barrier, as lipid constituents of topical products may act as a mechanical barrier and augment synthesis of barrier lipids. Natural oils are applied topically as part of a traditional oil massage to neonates in many developing countries. We sought to identify inexpensive, safe, vegetable oils available in developing countries that improved epidermal barrier function. The impact of oils on mouse epidermal barrier function (rate of transepidermal water loss over time following acute barrier disruption by tape-stripping) and ultrastructure was determined. A single application of sunflower seed oil significantly accelerated skin barrier recovery within 1 h; the effect was sustained 5 h after application. In contrast, the other vegetable oils tested (mustard, olive and soybean oils) all significantly delayed recovery of barrier function compared with control- or Aquaphor-treated skin. Twice-daily applications of mustard oil for 7 d resulted in sustained delay of barrier recovery. Moreover, adverse ultrastructural changes were seen under transmission electron microscopy in keratin intermediate filament, mitochondrial, nuclear, and nuclear envelope structure following a single application of mustard oil. CONCLUSION: Our data suggest that topical application of linoleate-enriched oil such as sunflower seed oil might enhance skin barrier function and improve outcome in neonates with compromised barrier function. Mustard oil, used routinely in newborn care throughout South Asia, has toxic effects on the epidermal barrier that warrant further investigation.

Attenuation coefficient and sound speed in human myometrium and uterine fibroid tumors.

OBJECTIVE: To develop a noninvasive method for treatment of uterine fibroid tumors using high-intensity focused ultrasound. Optimal high-intensity focused ultrasound treatment would be dependent on quantitative information about ultrasonic tissue characteristics. METHODS: Ultrasonic attenuation and the sound speed of fresh human fibroid tumors and myometrium were measured as a function of frequency (1-3 MHz) by using a pulse transmission technique before and after in vitro high-intensity focused ultrasound treatment (3.5 MHz at an intensity of 2,000 W/cm2). RESULTS: The ranges of the attenuation coefficients, before and after high-intensity focused ultrasound treatment, were 0.9 to 2.2 and 1.8 to 3.9 dB/cm2, respectively, for fibroid tumors and 0.5 to 1.6 and 1.7 to 3.3 dB/cm2, respectively, for myometrium. Although the sound speed appeared to be independent of frequency (1,611 to 1,616 m/s at 1 to 3 MHz) in both types of tissues, a slight increase of approximately 4 to 14 m/s was observed after high-intensity focused ultrasound treatment. CONCLUSIONS: The results of this study represent our first reported values of the attenuation coefficient and sound speed in fibroid tumors and myometrium before and after high-intensity focused ultrasound treatment.

Real-time visualization of high-intensity focused ultrasound treatment using ultrasound imaging.

High-intensity focused ultrasound (HIFU) and conventional B-mode ultrasound (US) imaging were synchronized to develop a system for real-time visualization of HIFU treatment. The system was tested in vivo in pig liver. The HIFU application resulted in the appearance of a hyperechoic spot at the focus that faded gradually after cessation of HIFU exposure. The duration of HIFU exposure needed for a hyperechoic spot to appear, was inversely related to the HIFU intensity. The threshold intensity required to produce a hyperechoic spot in liver in < 1 s was 970 W/cm(2), in situ. At this HIFU dose, no immediate cellular damage was observed, providing a potential for pretreatment targeting. The real-time visualization method was used in hemostasis of actively bleeding internal pelvic vessels, allowing targeting and monitoring of successful treatment. Real-time US imaging may provide a useful tool for image-guided HIFU therapy.

Impaired survival of bone marrow hematopoietic progenitor cells in cyclic neutropenia.

Cyclic neutropenia (CN) is a congenital hematopoietic disorder characterized by remarkably regular oscillations of blood neutrophils from near normal to extremely low levels at 21-day intervals. Recurring episodes of severe neutropenia lead to repetitive and sometimes life-threatening infections. To investigate the cellular mechanism of CN, the ultrastructure and the proliferative and survival characteristics of bone marrow-derived CD34(+) early progenitors, CD33(+)/CD34(-) myeloid progenitors, and CD15(+) neutrophil precursors from CN patients and healthy volunteers were studied. The ultrastructural studies showed profound apoptotic features in bone marrow progenitor cells in CN. Colony-forming assays demonstrated a 75% decrease in the number of early myeloid-committed colonies compared with controls. Long-term culture-initiating cell assays demonstrated a 6-fold increase in production of primitive progenitor cells in CN. To determine whether accelerated apoptosis might account for the underproduction of myeloid progenitors, the hematopoietic subpopulations were labeled with fluorescein isothiocyanate-annexin V and analyzed by flow cytometry. Short-term culture of CN cells resulted in apoptosis of approximately 65% of CD34(+) cells, 80% of CD33(+)/CD34(-) cells, and more than 70% of CD15(+) cells, as compared with 20%, 7%, and 15% apoptosis in respective control subpopulations. Evidence of accelerated apoptosis of bone marrow progenitor cells was observed in all 8 patients participating in the study, regardless of the stage in the CN cycle in which bone marrow aspirations were obtained. Granulocyte colony-stimulating factor therapy of CN patients significantly improved survival of bone marrow progenitor cells. These data indicate that ineffective production of neutrophils is due to accelerated apoptosis of bone marrow myeloid progenitor cells in CN.

Fas (CD95) induces alveolar epithelial cell apoptosis in vivo: implications for acute pulmonary inflammation.

Activation of the Fas/FasL system induces apoptosis of susceptible cells, but may also lead to nuclear factor kappaB activation. Our goal was to determine whether local Fas activation produces acute lung injury by inducing alveolar epithelial cell apoptosis and by generating local inflammatory responses. Normal mice (C57BL/6) and mice deficient in Fas (lpr) were treated by intranasal instillation of the Fas-activating monoclonal antibody (mAb) Jo2 or an irrelevant control mAb, and studied 6 or 24 hours later using bronchoalveolar lavage (BAL), histopathology, DNA nick-end-labeling assays, and electron microscopy. Normal mice treated with mAb Jo2 had significant increases in BAL protein at 6 hours, and BAL neutrophils at 24 hours, as compared to lpr mice and to mice treated with the irrelevant mAb. Neutrophil recruitment was preceded by increased mRNA expression for tumor necrosis factor-alpha, macrophage inflammatory protein-1alpha, macrophage inflammatory protein-2, macrophage chemotactic protein-1, and interleukin-6, but not interferon-gamma, transforming growth factor-ss, RANTES, eotaxin, or IP-10. Lung sections from Jo2-treated normal mice showed neutrophilic infiltrates, alveolar septal thickening, hemorrhage, and terminal dUTP nick-end-labeling-positive cells in the alveolar septae and airspaces. Type II pneumocyte apoptosis was confirmed by electron microscopy. Fas activation in vivo results in acute alveolar epithelial injury and lung inflammation, and may be important in the pathogenesis of acute lung injury.

Transcytosis of gastrointestinal epithelial cells by Escherichia coli K1.

Escherichia coli K1 is an important neonatal pathogen that is usually transferred from maternal to infant gastrointestinal tract at the time of parturition. Approximately 20% of neonates are colonized, and a proportion of colonized infants goes on to have systemic infection. Entry into the bloodstream from the gastrointestinal tract is hypothesized to occur via epithelial cell invasion. Invasion of multiple epithelial cell lines was studied using gentamicin protection assays and transcytosis of polarized monolayers. Electron microscopy was used to confirm cellular invasion. Cell lines used include two human gastrointestinal lines, Caco-2 and T84; a human respiratory cell line, A549; a human laryngeal cell line, HEp-2; and a canine kidney cell line, MDCK. A virulent E. coli K1 strain, RS218, readily invaded HEp-2, A549, and T84 cell lines in gentamicin protection assays, but was less invasive into MDCK and Caco-2 cells. RS218 also demonstrated transcytosis of both T84 and Caco-2 cells. Four clinical isolates of E. coli K1 demonstrated levels of transcytosis of T84 cells similar to RS218. Caco-2 invasiveness correlated with length of time in tissue culture with maximum invasiveness demonstrated at 11 d in culture, when cells were polarized and differentiated.

Counteracting effects of renal solutes on amyloid fibril formation by immunoglobulin light chains.

In primary (light chain-associated) amyloidosis, immunoglobulin light chains deposit as amyloid fibrils in vital organs, especially the kidney. Because the kidney contains high concentrations of urea that can destabilize light chains as well as solutes such as betaine and sorbitol that serve as protein stabilizers, we investigated the effects of these solutes on in vitro amyloid fibril formation and thermodynamic stability of light chains. Two recombinant light chain proteins, one amyloidogenic and the other nonamyloidogenic, were used as models. For both light chains, urea enhanced fibril formation by reducing the nucleation lag time and diminished protein thermodynamic stability. Conversely, betaine or sorbitol increased thermodynamic stability of the proteins and partially inhibited fibril formation. These solutes also counteracted urea-induced reduction in protein thermodynamic stability and accelerated fibril formation. Betaine was more effective than sorbitol. A model is presented to explain how the thermodynamic effects of the solutes on protein state equilibria can alter nucleation lag time and, hence, fibril formation kinetics. Our results provide evidence that renal solutes control thermodynamic and kinetic stability of light chains and thus may modulate amyloid fibril formation in the kidney.