Mammary tumors in Sprague Dawley rats induced by N-ethyl-N-nitrosourea for evaluating terahertz imaging of breast cancer.

Purpose: The objective of this study is to quantitatively evaluate terahertz (THz) imaging for differentiating cancerous from non-cancerous tissues in mammary tumors developed in response to injection of N-ethyl-N-nitrosourea (ENU) in Sprague Dawley rats. Approach: While previous studies have investigated the biology of mammary tumors of this model, the current work is the first study to employ an imaging modality to visualize these tumors. A pulsed THz imaging system is utilized to experimentally collect the time-domain reflection signals from each pixel of the rat's excised tumor. A statistical segmentation algorithm based on the expectation-maximization (EM) classification method is implemented to quantitatively assess the obtained THz images. The model classification of cancer is reported in terms of the receiver operating characteristic (ROC) curves and the areas under the curves. Results: The obtained low-power microscopic images of 17 ENU-rat tumor sections exhibited the presence of healthy connective tissue adjacent to cancerous tissue. The results also demonstrated that high reflection THz signals were received from cancerous compared with non-cancerous tissues. Decent tumor classification was achieved using the EM method with values ranging from 83% to 96% in fresh tissues and 89% to 96% in formalin-fixed paraffin-embedded tissues. Conclusions: The proposed ENU breast tumor model of Sprague Dawley rats showed a potential to obtain cancerous tissues, such as human breast tumors, adjacent to healthy tissues. The implemented EM classification algorithm quantitatively demonstrated the ability of THz imaging in differentiating cancerous from non-cancerous tissues.

Bariatric Radioembolization: A Pilot Study on Technical Feasibility and Safety in a Porcine Model.

PURPOSE: To evaluate feasibility of left gastric artery (LGA) yttrium-90 ((90)Y) radioembolization as potential treatment for obesity in a porcine model. MATERIALS AND METHODS: This study included 8 young female pigs (12-13 weeks, 21.8-28.1 kg). Six animals received infusions of (90)Y resin microspheres (46.3-105.1 MBq) into the main LGA and the gastric artery arising from the splenic artery. Animal weight and serum ghrelin were measured before treatment and weekly thereafter. Animals were euthanized 69-74 days after treatment, and histologic analyses of mucosal integrity and ghrelin immunoreactive cell density were performed. RESULTS: Superficial mucosal ulcerations < 3.0 cm(2) were noted in 5 of 6 treated animals. Ghrelin immunoreactive cell density was significantly lower in treated versus untreated animals in the stomach fundus (13.5 vs 34.8, P < .05) and stomach body (11.2 vs 19.8, P < .05). Treated animals gained less weight than untreated animals over the study duration (40.2 kg ± 5.4 vs 54.7 kg ± 6.5, P = .053). Average fundic parietal area (165 cm(2) vs 282 cm(2), P = .067) and average stomach weight (297.2 g vs 397.0 g, P = .067) were decreased in treated versus untreated animals. Trichrome staining revealed significantly more fibrosis in treatment animals compared with control animals (13.0 vs 8.6, P < .05). No significant differences were identified in plasma ghrelin concentrations (P = .24). CONCLUSIONS: LGA (90)Y radioembolization is promising as a potential treatment for obesity. A larger preclinical study is needed to evaluate the safety and efficacy of this procedure further.

The effects of differences in pspA alleles and capsular types on the resistance of Streptococcus pneumoniae to killing by apolactoferrin.

Pneumococcal surface protein A (PspA) is the only pneumococcal surface protein known to strongly bind lactoferrin on the bacterial surface. In the absence of PspA Streptococcus pneumoniae becomes more susceptible to killing by human apolactoferrin (apo-hLf), the iron-free form of lactoferrin. In the present study we examined diverse strains of S. pneumoniae that differed by 2 logs in their susceptibility to apo-hLf. Among these strains, the amount of apo-hLf that bound to cell surface PspA correlated directly with the resistance of the strain to killing by apo-hLf. Moreover examination of different pspA alleles on shared genetic backgrounds revealed that those PspAs that bound more lactoferrin conferred greater resistance to killing by apo-hLf. The effects of capsule on killing of pneumococci by apo-hLf were generally small, but on one genetic background, however, the lack of capsule was associated with 4-times as much apo-hLf binding and 30-times more resistance to killing by apo-hLf. Overall these finding strongly support the hypothesis that most of the variation in the ability of apo-hLf is dependent on the variation in the binding of apo-hLf to surface PspA and this binding is dependent on variation in PspA as well as variation in capsule which may enhance killing by reducing the binding of apo-hLf to PspA.

How Sensitive Is the Upper Gastrointestinal Tract to 90Y Radioembolization? A Histologic and Dosimetric Analysis in a Porcine Model.

In 90Y radioembolization, nontarget embolization to the stomach or small bowel can result in gastrointestinal injury, a rare but difficult to manage clinical complication. However, dosimetric thresholds for toxicity to these tissues from radioembolization have never been evaluated in a controlled setting. We performed an analysis of the effect of 90Y radioembolization in a porcine model at different absorbed-dose endpoints. METHODS: Six female pigs underwent transfemoral angiography and infusion of 90Y-resin microspheres into arteries supplying part of the gastric wall. Esophagogastroduodenoscopy was performed after 4 wk to assess interim gastrointestinal health. Animals were monitored for side effects for 9 wk after 90Y infusion, after which they were euthanized and their upper gastrointestinal tracts were excised for analysis. Histologic sections were used to map microsphere location, and a microdosimetric evaluation was performed to determine the absorbed-dose profile within the gastrointestinal wall. RESULTS: 90Y radioembolization dosages from 46.3 to 105.1 MBq were infused, resulting in average absorbed doses of between 35.5 and 91.9 Gy to the gastric wall. No animal exhibited any signs of pain or gastrointestinal distress through the duration of the study. Excised tissue showed 1-2 small (<3.0 cm2) healed or healing superficial gastric lesions in 5 of 6 animals. Histologic analysis demonstrated that lesion location was superficial to areas of abnormally high microsphere deposition. An analysis of microsphere deposition patterns within the gastrointestinal wall indicated a high preference for submucosal deposition. Dosimetric evaluation at the luminal mucosa performed on the basis of microscopic microsphere distribution confirmed that 90Y dosimetry techniques conventionally used in hepatic dosimetry provide a first-order estimate of absorbed dose. CONCLUSION: The upper gastrointestinal tract may be less sensitive to 90Y radioembolization than previously thought. Lack of charged-particle equilibrium at the luminal mucosa may contribute to decreased toxicity of 90Y radioembolization compared with external-beam radiation therapy in gastrointestinal tissue. Clinical examples of injury from 90Y nontarget embolization have likely resulted from relatively large 90Y activities being deposited in small tissue volumes, resulting in absorbed doses in excess of 100 Gy.

Use of CPR in hemorrhagic shock, a dog model.

INTRODUCTION: Cardiopulmonary resuscitation was designed for sudden cardiac events usually triggered by thrombotic phenomena. Despite this, it is routinely used in trauma resuscitations as per the American Heart guidelines. There is no data supporting the use of chest compressions in hemorrhagic shock. An evidence-based cardiopulmonary resuscitation (CPR) protocol has been developed for dogs. We sought to determine the effects and outcomes of chest compressions in hemorrhagic shock in a canine model. METHODS: Eighteen dogs were randomized to three treatment groups-chest compressions only after hemorrhagic shock (CPR), CPR with fluid resuscitation after hemorrhagic shock (CPR + FLU), and fluid resuscitation alone after hemorrhagic shock (FLU). Under anesthesia, dogs were hemorrhaged until pulse was lost; they were maintained pulseless for 30 minutes and then resuscitated over 20 minutes. Vital signs and laboratory values were recorded at determined intervals. Echocardiography was performed throughout the study. Upon termination of the study, kidney, liver, heart, and brain tissue histology was evaluated for end organ damage. Statistical significance was p < 0.05 with a Bonferroni correction for multiple comparisons. RESULTS: Blood loss and mean time to loss of pulse were similar between the groups. Dogs in the CPR group had significantly lower mean arterial pressure and higher pulse at all points compared to CPR + FLU and FLU (p < 0.05). Ejection fraction was lower in the CPR group at 5 and 10 minutes compared to the other groups (p < 0.05). Vital signs and laboratory results between CPR + FLU and FLU were equivalent. Two of six dogs in the CPR group died, while no dogs died in the CPR + FLU or FLU groups. Dogs in the CPR group were found to have more episodes of end organ damage. CONCLUSION: There was no benefit to chest compressions in the hypovolemic animals. Chest compressions in addition to fluid did not reverse signs of shock better than fluid alone. Further research is needed to define if there is a role of CPR in the trauma patient with hemorrhagic shock.

An Incidence of Pseudopregnancy Associated with the Social Enrichment of Rabbits (Oryctolagus cuniculi).

Here we describe a case of pseudopregnancy in a New Zealand White rabbit as a result of pair housing with an aggressive conspecific. Clinical signs included fur pulling and nest building that developed shortly after separation from the aggressor. An ovariohysterectomy was performed, and histopathologic findings support the diagnosis of pseudopregnancy. When introducing adult female rabbits to pair housing, stable pairs may be difficult to achieve because of the dominance-associated behavior that can occur as hierarchal relationships are formed. Does that are pair-housed after puberty should be monitored for aggressive behavior.

Exposure to chlorine dioxide gas for 4 hours renders Syphacia ova nonviable.

The purpose of our study was to evaluate the efficacy of chlorine dioxide gas for environmental decontamination of Syphacia spp. ova. We collected Syphacia ova by perianal cellophane tape impression of pinworm-infected mice. Tapes with attached ova were exposed to chlorine dioxide gas for 1, 2, 3, or 4 h. After gas exposure, ova were incubated in hatching medium for 6 h to promote hatching. For controls, tapes with attached ova were maintained at room temperature for 1, 2, 3, and 4 h without exposure to chlorine dioxide gas and similarly incubated in hatch medium for 6 h. Ova viability after incubation was assessed by microscopic examination. Exposure to chlorine dioxide gas for 4 h rendered 100% of Syphacia spp. ova nonviable. Conversely, only 17% of ova on the 4-h control slide were nonviable. Other times of exposure to chlorine dioxide gas resulted in variable effectiveness. These data suggest that exposure to chlorine dioxide gas for at least 4 h is effective for surface decontamination of Syphacia spp. ova.

Modified opsonization, phagocytosis, and killing assays to measure potentially protective antibodies against pneumococcal surface protein A.

The standard opsonophagocytosis killing assay (OPKA) for antibodies to pneumococcal capsular polysaccharide was modified to permit an evaluation of the protection-mediating antibodies to pneumococcal surface protein A (PspA). We found that by increasing the incubation time with the complement and phagocytes from 45 min to 75 min, the protective activity was readily detected. In another modification, we used a capsule type 2 target strain that expressed PspA but not pneumococcal surface protein C (PspC). With these modifications separately or in combination, rabbit antisera to the recombinant α-helical or proline-rich domains of PspA mediated >50% killing of the target strain. The ability of normal human sera to mediate the killing of pneumococci in this modified OPKA correlated with their levels of antibodies to PspA and their ability to protect mice against fatal infection with a type 3 strain. Passive protection of mice against pneumococci and killing in the modified OPKA were lost when normal human sera were adsorbed with recombinant PspA (rPspA) on Sepharose, thus supporting the potential utility of the modified OPKA to detect protective antibodies to PspA. In the standard OPKA, monoclonal antibodies to PspA were strongly protective in the presence of subprotective amounts of anti-capsule. Thus, the currently established high-throughput OPKA for antibodies to capsule could be modified in one of two ways to permit an evaluation of the opsonic efficacy of antibodies to PspA.

Effect of local anesthetic on microorganisms in a murine model of surgical site infection.

BACKGROUND: Surgical site infections are common, with an incidence of 1.5% to 5% for all types of surgery. In vitro studies suggest an antimicrobial effect of local anesthetic. We hypothesized that subcutaneous infiltration of local anesthetic before surgical incision would reduce the incidence of postoperative wound infection. METHODS: In a wound infection model using 4- to 6-week-old female mice, Staphylococcus aureus and Escherichia coli were inoculated in surgical wounds infiltrated with local anesthetic or saline. On day 5, the mice were killed and tissues were evaluated for viable bacterial numbers, presence of bacteria histologically, and degree of inflammation on a scale of 0 to 3 based on number and types of inflammatory cells and presence of necrosis. RESULTS: A one-way between-subjects analysis of variance with Tukey honestly significant difference post hoc comparisons showed no statistically significant difference in the degree of inflammation in mice infiltrated with lidocaine, lidocaine mixed with bupivacaine, or saline (p = 0.994, p = 0.337, and p = 0.792, respectively). A Tukey honestly significant difference post hoc analysis demonstrated that the saline (p = 0.038) and lidocaine mixed with bupivacaine (p = 0.006) had significantly lower degrees of inflammation than did the lidocaine group. A Bonferroni post hoc test demonstrated that those in the lidocaine (p = 0.003) and lidocaine mixed with bupivacaine (p = 0.008) groups had significantly higher inflammation than those in the saline group after controlling for the condition of the inocula. CONCLUSIONS: Infiltrate, whether saline, lidocaine, or lidocaine mixed with Marcaine, did not result in significantly different bacterial presence or higher degree of inflammation when controlling for experimental condition of bacterial inocula. Thus, subcutaneous infiltration of local anesthetic before a surgical incision is made does not reduce the incidence of bacterial growth or influence the degree of inflammation which alters infection rates.

The proline-rich region of pneumococcal surface proteins A and C contains surface-accessible epitopes common to all pneumococci and elicits antibody-mediated protection against sepsis.

Pneumococcal surface protein A (PspA) and PspC of Streptococcus pneumoniae are surface virulence proteins that interfere with complement deposition and elicit protective immune responses. The C-terminal halves of PspA and PspC have some structural similarity and contain highly cross-reactive proline-rich (PR) regions. In many PR regions of PspA and PspC, there exists an almost invariant nonproline block (NPB) of about 33 amino acids. Neither the PR regions nor their NPB exhibit the alpha-helical structure characteristic of much of the protection-eliciting N-terminal portions of PspA and PspC. Prior studies of PspA and PspC as immunogens focused primarily on the alpha-helical regions of these molecules that lack the PR and NPB regions. This report shows that immunization with recombinant PR (rPR) molecules and passive immunization with monoclonal antibodies reactive with either NPB or PR epitopes are protective against infection in mice. PR regions of both PspA and PspC were antibody accessible on the pneumococcal surface. Our results indicate that while PspA could serve as a target of these protective antibodies in invasive infections, PspC might not. When antibody responses to rPR immunogens were evaluated by using flow cytometry to measure antibody binding to live pneumococci, it was observed that the mice that survived subsequent challenge produced significantly higher levels of antibodies reactive with exposed PR epitopes than the mice that became moribund. Due to their conservation and cross-reactivity, the PR regions and NPB regions represent potential vaccine targets capable of eliciting cross-protection immunity against pneumococcal infection.

Unique long terminal repeat and surface glycoprotein gene sequences of feline leukemia virus as determinants of disease outcome.

The outcome of feline leukemia virus (FeLV) infection in nature is variable, including malignant, proliferative, and degenerative disorders. The determinants of disease outcome are not well understood but are thought to include viral, host, and environmental factors. In particular, genetic variations in the FeLV long terminal repeat (LTR) and SU gene have been linked to disease outcome. FeLV-945 was previously identified as a natural isolate predominant in non-T-cell neoplastic and nonneoplastic diseases in a geographic cohort. The FeLV-945 LTR was shown to contain unique repeat elements, including a 21-bp triplication downstream of the enhancer. The FeLV-945 SU gene was shown to encode mutational changes in functional domains of the protein. The present study details the outcomes of infection with recombinant FeLVs in which the LTR and envelope (env) gene of FeLV-945, or the LTR only, was substituted for homologous sequences in a horizontally transmissible prototype isolate, FeLV-A/61E. The results showed that the FeLV-945 LTR determined the kinetics of disease. Substitution of the FeLV-945 LTR into FeLV-A/61E resulted in a significantly more rapid disease onset but did not alter the tumorigenic spectrum. In contrast, substitution of both the FeLV-945 LTR and env gene changed the disease outcome entirely. Further, the impact of FeLV-945 env on the disease outcome was dependent on the route of inoculation. Since the TM genes of FeLV-945 and FeLV-A/61E are nearly identical but the SU genes differ significantly, FeLV-945 SU is implicated in the outcome. These findings identify the FeLV-945 LTR and SU gene as determinants of disease.

Subtle mutational changes in the SU protein of a natural feline leukemia virus subgroup A isolate alter disease spectrum.

FeLV-945 is a representative isolate of the natural feline leukemia virus (FeLV) variant predominant in non-T-cell malignant, proliferative, and degenerative diseases in a geographic cohort. The FeLV-945 surface glycoprotein (SU) is closely related to natural horizontally transmissible FeLV subgroup A (FeLV-A) but was found to differ from a prototype to a larger extent than the members of FeLV-A differ among themselves. The sequence differences included point mutations restricted largely to the functional domains of SU, i.e., VRA, VRB, and PRR. Despite the sequence differences in these critical domains, measurements of receptor utilization, including host range and superinfection interference, confirmed the assignment of FeLV-945 to subgroup A. Other proviruses isolated from the cohort contained similar sequence hallmarks and were assigned to FeLV subgroup A. A provirus from cat 1046 contained a histidine-to-proline change at SU residue 6 within an SPHQ motif that was previously identified as a critical mediator of fusion events during virus entry. The 1046 pseudotype virus entered cells only in the presence of the soluble cofactor FeLIX provided in trans, but it retained an ecotropic host range even in the presence of FeLIX. The mutational changes in FeLV-945 were shown to confer significant functional differences compared to prototype FeLV-A viruses. The substitution of FeLV-945 envelope gene sequences for FeLV-A/61E sequences conferred a small but statistically significant replicative advantage in some feline cells. Moreover, substitution of the unique FeLV-945 long terminal repeat and envelope gene for those of FeLV-A/61E altered the disease spectrum entirely, from a thymic lymphoma of a T-cell origin to an as yet uncharacterized multicentric lymphoma that did not contain T cells.