Effects of dietary supplementation of formaldehyde and crystalline amino acids on gut microbial composition of nursery pigs.

Formaldehyde-based feed additives are approved in the US for Salmonella control and reducing bacterial contamination in animal feed. However, we hypothesize formaldehyde inclusion in swine diets may influence gut microbial composition due to its antimicrobial properties which might negatively influence microbial populations and pig growth performance. Also, formaldehyde inclusion in diets is known to reduce the dietary availability of amino acids. Therefore, our study was conducted to characterize if the effects of feed formaldehyde-treatment are due to influences on microbial population or diet amino acid (AA) sources. Dietary treatments were arranged in a (2 × 2) + 1 factorial with formaldehyde treatment (none vs. 1000 ppm formaldehyde) and crystalline AA inclusion (low vs. high) with deficient AA content plus a positive control diet to contain adequate AA content without dietary formaldehyde. Treating diets with formaldehyde reduced growth rate (P = 0.001) while the AA inclusion had no evidence of impact. Formaldehyde reduced feed bacterial content and altered fecal microbial communities (P < 0.05). Therefore, we conclude that the negative influence on growth was due to the impact on the fecal microbial community. Implications are that strategies for feed pathogen control need to take into account potential negative impacts on the gut microbial community.

Case Series Report of 66 Refractory Maintenance Patients Evaluating the Effectiveness of Topical Oxidizing Agents.

OBJECTIVE: To evaluate the clinical effects of a prescription tray application of hydrogen peroxide gel as an adjunct to frequent maintenance appointments for refractory periodontal patients. METHODS: Case series data were analyzed from 66 failing periodontal maintenance patients who had exhausted treatment options before using prescription trays with a 1.7% hydrogen peroxide gel once or twice daily for two-and-a-half to five years. Data included pocket probing depths (PPD), bleeding on probing (BOP), smoking status, and compliance with tray usage. Data were collected prior to tray usage and after tray delivery at six months, one year, and annual intervals. RESULTS: A clinical and statistical reduction in BOP was maintained over the length of the study (p ≤ 0.01). No differences were seen in patients who used trays two times or one time a day or in patients who smoked or did not smoke. The 1.7% peroxide delivered via a prescription tray was most effective in shallow pockets. Aggregate PPD distribution did not change significantly. Of clinical relevance, only one tooth out of 1,745 teeth studied was lost due to periodontal disease during the study period. CONCLUSION: Prescription tray application of peroxide gel, as an adjunct to frequent periodontal maintenance appointments for refractory patients, demonstrated significant reductions in BOP'for smokers and non-smokers who used tray delivery once or twice a day.

Cosmesis and satisfaction after breast-conserving surgery correlates with the percentage of breast volume excised.

BACKGROUND: The cosmetic outcome after breast-conserving surgery correlates with psychosocial adjustment. Previous studies have shown that, among other factors, cosmesis is dependent on breast size and weight of the wide local excision specimen. This study assessed cosmetic outcome relative to a combination of these factors-the estimated percentage of breast volume excised (EPBVE). METHODS: The study group consisted of 151 women who had undergone breast-conserving surgery. All had previously completed a patient satisfaction questionnaire and an independent panel had assessed cosmetic outcome. Their mammograms were reviewed and breast volume was estimated. A validation series showed cone volume on the oblique mammogram to predict true breast weight most accurately (r = 0.93). RESULTS: Both subjective cosmetic assessment and patient satisfaction correlated strongly with EPBVE. Overall, when the EPBVE was below 10 per cent, 83.5 per cent of patients were very satisfied with their appearance and only 3.1 per cent were not satisfied, compared with 37.0 and 16.7 per cent respectively if the EPBVE was more than 10 per cent. However, tumour location in the breast was also an important factor; cosmetic outcome was worse for medial tumours. CONCLUSION: The percentage of breast volume excised was an important determinant of cosmesis and patient satisfaction after breast-conserving surgery. Calculating the EPBVE before surgery may help select women in whom breast reshaping or volume replacement should be considered.

cAMP-mediated suppression of a Th1 clone associated with an alteration of the intracellular redox environment.

We have shown previously that the elevation of intracellular cAMP in antigen or anti-CD3-activated murine Th1 clones in the absence of antigen inhibits antigen-induced proliferation and the production of IL-2 by H2O2-mediated oxidation of p56lck and inhibits antigen-induced production of interferon-gamma by the induction of intracellular nitric oxide. Moreover, activated Th1 clones are resistant to cAMP-induced suppression. These results suggest that the immunosuppression of Th1 cells mediated by elevated intracellular cAMP is associated with an alteration in the intracellular oxidation/reduction environment. Here we report that the culture of an antigen or anti-CD3-activated murine Th1 clone with the adenylcyclase agonist forskolin (FSK) in the absence of antigen reduces the activity of intracellular catalase, and diminishes levels of intracellular reduced glutathione (GSH). Resting cells resistant to cAMP-induced suppression have higher intracellular GSH levels than antigen-activated cells susceptible to cAMP-induced suppression. The results provide further evidence that cAMP-induced suppression of Th1 clones is mediated by profound alterations in the intracellular redox environment and may be used to selectively inactivate Th1 cells activated by antigen.

Differential regulation of T cell receptor-mediated Th1 cell IFN-gamma production and proliferation by divergent cAMP-mediated redox pathways.

Culture of an H-2(s)-restricted, bovine myelin basic protein (BMBP)-specific murine Th1 clone with the adenyl cyclase agonist forskolin (FSK) or isobutylmethylxanthine (IBMX), an inhibitor of cAMP catabolism, before culture with anti-CD3 or BMBP and antigen-presenting cells (APC) suppressed antigen or anti-CD3-induced proliferation and production of interferon-gamma (IFN-gamma). Other H-2(s)-derived or H-2(b)-derived clones specific for BMBP or keyhole limpet hemocyanin (KLH) were similarly affected. FSK did not affect the expression of CD4 or the T cell receptor (TCR) but did diminish levels of the phosphorylated (activated) mitogen-activated protein (MAP) kinases early response kinase-1 (ERK-1) and ERK-2. Immunoblotting of lysates from an FSK-treated Th1 clone with antibodies to a carboxy-terminal epitope of p56(lck), a signal transduction enzyme upstream from ERK-1 and ERK2, did not detect p56(lck) unless the lysates were reduced prior to electrophoresis. Immunoblotting of nonreduced lysates with antibodies to an amino-terminal epitope demonstrated p56(lck) with a lower apparent molecular weight, characteristic of oxidized proteins. Reduction restored the detection of p56(lck) by anticarboxy-terminal p56(lck) and to mobilities indistinguishable from controls detected by the antiamino-terminal p56(lck). N-acetylcysteine or catalase prevented FSK-induced suppression of antigen-induced proliferation and the loss of carboxy-terminal epitopes of p56(lck). An inhibitor of cAMP-dependent protein kinase A (PKA) or nitric oxide synthase (NOS) did not affect FSK-induced inhibition of antigen-induced proliferation. In contrast, inhibitors of PKA or NOS, but not catalase, prevented FSK-induced suppression of IFN-gamma production. Moreover, immunoblots of lysates precipitated with anti-p56(lck), phosphotyrosine, or CD4 demonstrated that in FSK-treated, anti-CD3-stimulated cells, p56(lck) is not associated with CD4 zeta chain, nor is p56(lck) or zeta chain phosphorylated. In vitro kinase assays demonstrated that p56(lck) from FSK-treated cells does not have kinase activity. Taken together, the results suggest that an elevation of intracellular cAMP (in the absence of antigen) creates an oxidative environment that oxidizes and inactivates p56(lck) by an H(2)O(2)-dependent, PKA-independent mechanism and inhibits the production of IFN-gamma by an NO, PKA-dependent mechanism. Thus, antigen-induced proliferation and IFN-gamma production in a Th1 clone are controlled separately by different cAMP-dependent, redox-based mechanisms.

Chronic haematoma as a late complication of cosmetic breast augmentation.

To our knowledge, a chronic expanding haematoma has not been reported in a patient who received silicone gel breast prostheses for augmentation and never before at an interval as long as 12 years, as in our case. There have been occasional reports of chronic haematoma as complication of implantation of silicone breast prostheses after mastectomy for cancer, presenting 2.5-8 years later.

The immunohistochemical expression of desmoplakin and its role in vivo in the progression and metastasis of breast cancer.

Desmoplakin (DP) is a protein located at the inner plaque of desmosomes where it associates with the desmosomal cadherins to form a cell adhesion complex. Reduced expression of DP has been correlated with the progression of several cancers, but its role in in vivo breast cancer is yet to be established. The aim of this present paper was to determine the immunohistochemical (IHC) expression of DP in breast cancer specimens (n = 75) in comparison with ductal carcinoma in situ (DCIS) (n = 26), tumour associated normal (n = 29) and normal breast tissue (n = 7). DP expression was correlated with that of desmosomal cadherin, Desmoglein 2 (Dsg2) and other clinical and IHC prognostic markers. DP staining occurred at the sub-plasma membrane level. There was no significant correlation between the level of DP (as assessed by the H-score) and that of Dsg. Significantly stronger staining was demonstrated in normal breast tissue and well differentiated tumours compared with more moderately or poorly differentiated tumours (P = 0.04). A significant inverse correlation was seen between DP staining and tumour size (P = 0.01). In a limited series of 8 cases, primary tumours demonstrated significantly stronger staining than the matched metastatic lymph nodes (P = 0.046). Of all the IHC markers examined, only Ki-67 showed a significant inverse relationship with DP staining (P = 0.01). In summary, the data suggest that loss of DP may be of potential importance in progression of breast cancer in vivo from normal, DCIS, well differentiated through to poorly differentiated, large tumours. In addition, this loss may be associated with metastasis.

The National Breast Referral Guidelines have cut down inappropriate referrals in the under 50s.

AIMS: When the National Breast Referral Guidelines were applied to our local GPs letters immediately prior to their release in January 1996, it was shown that on the basis of the GPs own conclusions that 29% of symptomatic women could have been managed initially by their own GP without missing any carcinomas. We conducted this study at the Rapid Access Breast Clinic at the University Hospital of Wales to determine if the breast referral practices of local GPs have altered due to the breast referral guidelines. METHODS: We studied 2332 referrals from the inception of the Rapid Access Clinic in May 1995 to the issue of the guidelines, and 2421 referrals from May 1996 to the end of the year. Random samples of 600 patients were drawn from each year and the referral letters were scored as within or outside the guidelines. Family history patients were excluded. RESULTS: There was an 11% fall in referrals outside the guidelines in the under 50s (chi-squared=<0.001) but the 7% fall in the over 50s was not significant. CONCLUSIONS: The Breast Referral Guidelines seem to have been effective in reducing the higher level of inappropriate referrals in younger patients at less risk of carcinoma.

Is there a role for surgery in the treatment of mastalgia?

Breast pain is a common condition affecting most women at some stage in their reproductive life. Mastalgia is resistant to treatment in 6% of cyclical and 26% non-cyclical patients. Surgery is not widely used to treat this condition and only considered in patients with severe mastalgia resistant to medication. The aims of this study were to audit the efficacy of surgery in severe treatment resistant mastalgia and to assess patient satisfaction following surgery. This is a retrospective review of the medical records of all patients seen in mastalgia clinic in the University Hospital of Wales, Cardiff since 1973. A postal questionnaire was distributed to all patients who had undergone surgery. Results showed that of the 1054 patients seen in mastalgia clinic, 12 (1.2%) had undergone surgery. Surgery included 8 subcutaneous mastectomies with implants (3 bilateral, 5 unilateral), 1 bilateral simple mastectomy and 3 quadrantectomies (1 having a further simple mastectomy). The median duration of symptoms was 6.5 years (range 2-16 years). Five patients (50%) were pain free following surgery, 3 developed capsular contractures and 2 wound infections with dehiscence. Pain persisted in both patients undergoing quadrantectomy. We conclude that surgery for mastalgia should only be considered in a minority of patients. Patients should be informed of possible complications inherent of reconstructive surgery and warned that in 50% cases their pain will not be improved.

Senior surgeons and radiologists should assess emergency patients on presentation: a prospective randomized controlled trial.

The objective of this study was to determine whether abdominal ultrasound and plain film radiography (evaluated by senior radiologists) substantially enhances senior surgical assessment in reducing surgical admissions. A randomized controlled trial was carried out in a teaching hospital; ninety-five emergency referrals to general surgery (mean age 51 years, 37% males), presenting with intrabdominal disturbances not requiring immediate surgery, were selected for study. The major outcome measure was the number of patients not admitted as in-patients. Thirty per cent of the intervention group and 10% of the controls did not need admission (95% confidence interval on this 29% difference, 12-38%). There were no important differences in the mean waiting time in casualty (26 min vs. 25 min), length of admission (4.1-5.9 days), surgical intervention (seven vs. 11 patients), readmission with similar pathology within 3 months (seven vs. three patients), and mortality as an in-patient or within 3 months of discharge. Abdominal ultrasound and plain film radiography, assessed by senior radiologists, enhances senior surgical assessment in reducing unnecessary surgical admissions.

Evaluation of general practitioner referrals to a specialist breast clinic according to the UK national guidelines.

The recently published national guidelines to general practitioners for the referral of patients with breast problems were retrospectively applied to letters of all patients attending the Rapid Access Breast Clinic at the University Hospital of Wales. The patients have all had diagnostic investigations performed at the initial visit with a multidisciplinary review of results and provision of a management plan prior to the subsequent visit. Since its inception in May 1995 until the end of the year when the guidelines were published, 2332 new patients had been seen. Overall, 29% of patients with benign breast disease would not have been referred if the guidelines had been strictly followed. Of the 147 symptomatic carcinomas diagnosed from general practitioner referrals (6.3% of total referrals), no invasive cancers would have been missed. One patient with incidental detection of ductal carcinoma in situ (DCIS) in the asymptomatic contralateral breast would not have been referred. Referral for pain without a discrete lump constituted 63% of the patients with a benign diagnosis who fell outside the guidelines. The guidelines also include comprehensive algorithms for the initial management of benign breast symptoms that do not require immediate referral by the general practitioners. Our general practitioners were significantly better at referring patients over 50 years old but the proportion of appropriate referrals were not related to the number referred by each practice. The present guidelines adequately cover referral for the diagnosis of malignant breast disease to a specialist, and may reduce the benign workload of breast clinics.

The role of nitric oxide in cancer. Improved methods for measurement of nitrite and nitrate by high-performance ion chromatography.

The short lifetime of nitric oxide (NO) in vivo impedes its quantitation directly; however, the determination of nitrite and nitrate ions as the end-products of NO oxidation has proven a more practical approach. High-performance ion chromatographic analysis of nitrite in biological fluids is hampered by the large amount of chloride ion (up to approximately 100 mmol/l) which results in insufficient peak resolution when utilizing conductimetric detection. Analysis of both anions in small sample volumes is also constrained by the need to minimise sample handling to avoid contamination by environmental nitrate. We report a means to remove Cl- ions from small sample volumes using Ag+ resin which facilitates quantitation of either nitrite and nitrate anions in biological samples, using silica or polymer based ion-exchange resins with conductimetric or electrochemical and spectrophotometric detection. Including a reversed-phase guard column before the anion-exchange guard and analytical column also greatly extends column lifetime.

Undetected gynaecological disorders in women with renal disease.

Women with chronic renal disease (CRD) who are on dialysis or have a functioning renal transplant are typically stoical in their attitude towards other health problems. We undertook a prospective study of 100 women with CRD to assess the prevalence of gynaecological disorders in this group of patients. Assessment included the measurement of follicle stimulating hormone, luteinizing hormone, prolactin and oestradiol concentrations, cervical cytology and a pelvic ultrasound scan. We found that gynaecological problems are highly prevalent and frequently unrecognized. Of these women, 58% had a menstrual disorder, with uncontrolled menorrhagia being a significant problem when it aggravated the chronic anaemia of renal disease, and 35% were menopausal, including seven women under the age of 40 years. Menopausal symptoms were undertreated. We identified a 14-fold increase in premature ovarian failure secondary to CRD and the use of cyclophosphamide therapy. In all, 22% of the women were subfertile and 10% had an abnormal smear, with cervical dyskariosis being significantly increased because of long-term immunosuppression. Contraceptive advice had often been absent or inappropriate. We conclude that formal gynaecological review should be routinely available for women with CRD.

The role of desmoglein 2 and E-cadherin in the invasion and motility of human breast cancer cells.

Loss of adhesion is a fundamental step in the metastatic cascade. Desmosomal cadherins, Desmoglein (Dsg) and Desmocollin (Dsc) are a novel group of adhesion molecules. Aims were to demonstrate expression of Dsg2 and E-cadherin in breast cancer cells and assess their role in invasion and motility. Immunofluorescence and Western blotting were used to demonstrate expression of Dsg2 and E-cadherin in 3 breast cancer cell lines (MDA MB 231, MCF7 and BT474). Functional studies included cell-cell aggregation, in vitro invasion and colloidal gold phagokinetic tracking assays. All 3 cell lines expressed Dsg2. MCF7 and BT474 cells were E-cadherin positive, MDA 231 was negative. Cell aggregation was reduced, in vitro invasion and motility were increased in Dsg2 or E-cadherin Mab pre-treated cells. Dsg2 present in breast cancer cells may act as a tumour suppressor molecule.

Elevation of intracellular cyclic AMP induces an anergic-like state in Th1 clones.

Because elevated intracellular cAMP suppresses T cell receptor (TCR)-mediated effector activity and/or proliferation in response to antigen but does not always affect IL-2-stimulated proliferation, the effects of cAMP on a T lymphocyte response to antigen resemble antigen-induced anergy. To test the hypothesis that elevated cAMP induces anergy in T lymphocytes, we have precultured murine Th1 clones responsive to porcine myelin basic protein (PMBP) with dibutyryl cyclic AMP (dbcAMP) or forskolin and subsequently removed the dbcAMP or forskolin and measured the proliferative response of the clones to antigen and antigen-presenting cells (APC) in the presence or absence of exogenously added interleukin-2 (IL-2). Cells precultured with dbcAMP or forskolin for 3 days did not proliferate or produce IL-2 in response to antigen and APC, but did proliferate to antigen and APC in the presence of IL-2. Cells that had not been stimulated recently with antigen/APC or IL-2 were not affected by dbcAMP, while cells stimulated recently with antigen/APC and IL-2 were susceptible to the anergizing effect of dbcAMP. These observations support the hypothesis that elevation in intracellular cAMP in antigen-activated Th1 clones, prior to subsequent culture with antigen, induces a state of anergy.

A 40-week male fertility study of the anticancer agent sulofenur (LY186641) administered orally to rats.

Sulofenur was evaluated for fertility effects on rats. Five-week old male rats (20/group) received 0, 5, 30, or 60 mg Sulofenur/kg/day for 14 weeks. Fertility was evaluated five times. Treated males were mated with untreated females at 10 weeks. Half the males from each group were necropsied after the 14-week treatment period and the remainder were mated four additional times during a 26-week posttreatment period. Following each mating, females were killed on gestation-day 20 and examined for evidence of pregnancy. Six weeks after mating trial 5, the remaining males were necropsied. Testes and epididymides were collected, weighed, and examined microscopically. All rats mated in each mating trial, and all control and low-dose animals were fertile. A significant reduction in fertility occurred in the middle- and high-dose males. This was consistent with testicular hypospermatogenesis in these groups. Fertility recovered in the high-dose group following cessation of treatment, but remained reduced in the middle-dose group. Preimplantation and postimplantation loss in mating trial 1 were higher in the middle- and high-dose groups. Abnormal fetuses were present in the high-dose group in mating trial 3, but not in mating trials 4 or 5. In conclusion, male rats given doses of 30 and 60 mg/kg/day of Sulofenur showed hypospermatogenesis and decreased fertility. Spermatogenesis and fertility recovered in the high-dose group. A dose of 5 mg/kg/day did not produce any effect on testes or fertility.

Skin pustules and nodules caused by Actinomyces viscosus in a horse.

A 12-year-old Arabian stallion was evaluated for a refractory skin problem of 2 years' duration. Skin nodules and pustules, which would rupture, had developed over the right lumbar muscles. Anaerobic culturing from the pustules yielded Actinomyces viscosus, and histologic examination of biopsy specimens revealed globular eosinophilic structures. Concomitant treatment with isoniazid (8 mg/kg of body weight, q 24 h for 8 weeks), trimethoprim-sulfadiazine (30 mg/kg, q 24 h for 8 weeks), and sodium iodide solution (66 mg/kg, every 1, 2, or 4 weeks, for 32 weeks) resolved the condition.

Perioperative stroke risk in 173 consecutive patients with a past history of stroke.

During an 8-year period ending in 1988, 173 consecutive patients with a history of previous cerebrovascular accident underwent general anesthesia for surgery. Five patients (2.9%) had documented postoperative cerebrovascular accidents from 3 to 21 days (mean, 12.2 days) after surgery. The risk of postoperative cerebrovascular accident did not correlate with age, sex, history of multiple cerebrovascular accidents, poststroke transient ischemic attacks, American Society for Anesthesia physical status, aspirin use, coronary artery disease, peripheral vascular disease, intraoperative blood pressure, time since previous cerebrovascular accident, or cause of previous cerebrovascular accident. Postoperative stroke was more common in patients given preoperative heparin sodium. We conclude that the risk of perioperative stroke is low (2.9%) but not easily predicted and that the risk continues beyond the first week of convalescence. Unlike myocardial infarction, cerebral reinfarction risk does not seem to depend on time since previous infarct.