Neonatal gut colonization by Staphylococcus aureus strains with certain adhesins and superantigens is negatively associated with subsequent development of atopic eczema.

BACKGROUND: Insufficient early immune stimulation may predispose to atopic disease. Staphylococcus aureus, a skin and gut colonizer, produces the B-cell mitogen protein A and T-cell-activating superantigens. Early gut colonization by S. aureus strains that possess the superantigens encoded by the enterotoxin gene (egc) cluster and elastin-binding protein is negatively associated with development of atopic eczema. OBJECTIVES: To investigate (i) whether these findings could be replicated in a second birth cohort, FARMFLORA, and (ii) whether nasal colonization by S. aureus also relates to subsequent atopic eczema development. METHODS: Faecal samples and nasal swabs from infants in the FARMFLORA birth cohort (n = 65) were cultured for S. aureus. Individual strains were distinguished by random amplified polymorphic DNA and assessed for adhesin and superantigen gene carriage by polymerase chain reaction. Atopic eczema at 18 months of age was related to nasal and gut S. aureus colonization patterns during the first 2 months of life (well before onset of eczema). RESULTS: Staphylococcus aureus colonization per se was unrelated to subsequent eczema development. However, gut S. aureus strains from the infants who subsequently developed atopic eczema less frequently carried the ebp gene, encoding elastin-binding protein, and superantigen genes encoded by egc, compared with strains from children who remained healthy. Nasal colonization by S. aureus was less clearly related to subsequent eczema development. CONCLUSIONS: The results precisely replicate our previous observations and may suggest that mucosal colonization by certain S. aureus strains provides immune stimulation that strengthens the epithelial barrier and counteracts the development of atopic eczema.

Short-term outcome after gluteus maximus myocutaneous flap reconstruction of the pelvic floor following extra-levator abdominoperineal excision of the rectum.

AIM: Extra-levator abdominoperineal excision (APE) of the rectum has been introduced with the aim of improving the oncological outcome of low rectal cancer. The procedure includes resection of the levator muscles en bloc with the mesorectum, leaving a larger perineal defect than after conventional APE. This study reports short-term outcome of gluteus maximus myocutaneous flap reconstruction on perineal wound healing. METHOD: Sixty-five patients were studied after extra-levator APE and a one-sided myocutaneous flap for a low or locally recurrent rectal cancer at the Karolinska University Hospital from January 2002 to December 2008. Fifty-nine had received neoadjuvant radio- or radiochemotherapy. All perineal complications occurring within 30 days after surgery were registered. In addition, the status of the perineal reconstruction at 6 months and 1 year after surgery was assessed based on medical records from outpatient visits. RESULTS: Twenty-seven (41.5%) patients had one or more perineal wound complications. A minor wound infection occurred in 15, while 12 had either a more severe infection with dehiscence or a pelvic abscess. The reconstruction was completely healed in 91% of the patients at 1 year. CONCLUSION: Although the vast majority of the perineal reconstructions were healed at 1 year, the short-term perineal wound complication rate of gluteus maximus flap reconstruction was high.

Effects of continuous elevated cortisol concentrations during oestrus on concentrations and patterns of progesterone, oestradiol and LH in the sow.

This study investigated the effect of continuous elevated cortisol concentrations during standing oestrus on time of ovulation and patterns of progesterone, oestradiol and luteinising hormone (LH) in sows. The elevation of cortisol concentrations was achieved through repeated intravenous injections of synthetic adrenocorticotropic hormone (ACTH) every 2 h for approximately 48 h, from the onset of the second standing oestrus after weaning. Treatment was terminated when ovulation was detected (monitored by transrectal ultrasonography every 4h) or when the sow had received a maximum of 24 injections. The dose of ACTH (2.5 microg/kg) was chosen to mimic the cortisol concentrations seen during mixing of unfamiliar sows. The sows (n=14) were surgically fitted with jugular vein catheters and randomly divided into a control (C group where only NaCl solution were injected) or an ACTH group. Blood samples were collected every 2 h. In parallel with the blood sampling, saliva samples for cortisol analyses were taken from eight sows before onset of treatment and from four of the sows during treatment. There was no difference in time from onset of standing oestrus to ovulation between the two groups. The interval between the peaks of oestradiol and LH to ovulation was prolonged in the ACTH group compared to the C group (p<0.05), with a tendency towards an earlier decline of oestradiol in the ACTH group. Cortisol and progesterone concentrations were significantly elevated during treatment in the ACTH group (p<0.001), with cortisol peak concentrations occurring between 40 and 80 min after each ACTH injection. Cortisol concentrations in saliva and plasma were highly correlated (p<0.001). In conclusion, elevated cortisol concentrations from the onset of standing oestrus increase progesterone concentrations and prolong the interval between oestradiol and LH peaks to ovulation, the latter possible due to an early decline in oestradiol concentrations and a change of the LH peak outline. The effect these hormonal changes have on reproductive performance need to be further investigated. Saliva samples might be a useful and non-invasive method to assess cortisol concentrations in sows.

Impact of exogenous ACTH during pro-oestrus on endocrine profile and oestrous cycle characteristics in sows.

Sows housed in freely moving groups have elevated cortisol levels until the rank order is established, which takes place within approximately 48 h. The aim of this investigation was to study the effect of repeated administration of synthetic adrenocorticotropic hormone (ACTH; Synacthen Depot), during the follicular phase (pro-oestrus) on oestrus, ovulation and endocrine parameters. Four multiparous sows were used. Follicular growth and ovulation were recorded by ultrasonography. The first oestrous cycle after weaning was used as control cycle. Onset of oestrus in the sow occurs 3-4 days after the time when plasma progesterone reaches a concentration of 8 nmol/l. The progesterone profile in the control cycle of the individual sow was used for estimation when the ACTH injections should start. In the third pro-oestrus ACTH (2.5 microg/kg) was given via an indwelling catheter every 2 h for 48 h. The sows were euthanased 4-6 days after onset of the third oestrus and the ovaries were examined. Cortisol levels were elevated during the treatment period (p < 0.05). The second cycle, in which the sows were injected with ACTH, was prolonged with 2.5 days compared with the control cycle (p < 0.05). The oestradiol pattern during oestrus was similar in the control and the treatment cycle in ovulating sows. Three sows had ovulated (fresh corpora lutea), but the ovaries contained additionally one or several luteinized follicles/cysts. In conclusion, ACTH administration during pro-oestrus caused a prolongation of the oestrous cycle and a disturbed follicular development.

Extended abdominoperineal resection with gluteus maximus flap reconstruction of the pelvic floor for rectal cancer.

BACKGROUND: Intraoperative tumour perforation, positive tumour margins, wound complications and local recurrence are frequent difficulties with conventional abdominoperineal resection (APR) for rectal cancer. An alternative technique is the extended posterior perineal approach with gluteus maximus flap reconstruction of the pelvic floor. The aim of this study was to report the technique and early experience of extended APR in a select cohort of patients. METHODS: The principles of operation are that the mesorectum is not dissected off the levator muscles, the perineal dissection is done in the prone position and the levator muscles are resected en bloc with the anus and lower rectum. The perineal defect is reconstructed with a gluteus maximus flap. Between 2001 and 2005, 28 patients with low rectal cancer were treated accordingly at the Karolinska Hospital. RESULTS: Two patients had ypT0 tumours, 20 ypT3 and six ypT4 tumours. Bowel perforation occurred in one, the circumferential resection margin (CRM) was positive in two, and four patients had local perineal wound complications. Two patients developed local recurrence after a median follow-up of 16 months. CONCLUSION: The extended posterior perineal approach with gluteus maximus flap reconstruction in APR has a low risk of bowel perforation, CRM involvement and local perineal wound complications. The rate of local recurrence may be lower than with conventional APR.

Characterisation of cells in regenerating cartilage from autotransplanted perichondrium. immunohistochemical expression of smooth-muscle actin, desmin, vimentin, and Ki-67.

Autotransplanted perichondrium from rib and ear sutured to the knee joints of 26 rabbits has been examined with immunohistochemistry and shows certain structural, functional, and proliferative characteristics of regenerating cartilage. Cryostat sections have been examined for the expression of smooth-muscle actin (SMA), desmin, vimentin, and Ki-67. In this rabbit model of perichondrial grafting SMA staining showed vivid vessel regeneration, particularly in the proliferating stage about two to three weeks after grafting, and no vessels in more mature parts one month or more after transplantation. Desmin staining showed expression and distribution similar to SMA. Vimentin staining shows the cytoskeleton of regenerating cartilaginous tissue and makes cellular borders apparent. Immunohistochemical expression of Ki-67 is constantly negative in perichondrial tissue from rib and ear before transplantation, clearly positive in the proliferative stage, but there is no expression in maturing cartilage. The study also shows that all human antibodies used are applicable in a rabbit model.

Clones of normal keratinocytes and a variety of simultaneously present epidermal neoplastic lesions contain a multitude of p53 gene mutations in a xeroderma pigmentosum patient.

A patient with xeroderma pigmentosum group C was extensively examined for mutations in the p53 gene in normal skin exposed to varying degrees of sunlight and in excisional biopsies of basal cell cancer, squamous cell cancer, and squamous cell dysplasia. Seventy-three samples were analyzed by microdissection of small cell clusters, followed by PCR and direct DNA sequencing. In skin taken from areas that most likely had never been exposed to the sun, no mutations were found. However, in skin exposed to the sun, we observed a multitude of mutations in the p53 gene. UV light-induced mutations were found in all types of lesions, as well as in clusters of morphologically normal epidermal cells. Twenty-nine distinct mutations were found in exons 5-8, all missense or nonsense, of which 27 (93%) were UV-specific C --> T or CC --> TT transitions at dipyrimidine sites of the nontranscribed strand. Two types of normal skin areas containing p53 mutations were observed: areas that stain strongly with p53 antibody (p53 patches) and those that do not stain. Because no silent or intron mutations were found in these cell clusters, the alterations in the p53 gene of morphologically normal cells are likely to have resulted in a selective growth advantage. The poor correlation between mutations and morphological phenotypes demonstrates that p53 mutations alone do not determine the phenotypes observed.

Binding of extracellular matrix proteins to the surface of Bacteroides spp.

The binding of fibronectin an vitronectin to 207 Bacteroides strains and the binding of collagen and sialoproteins to 55 Bacteroides strains were investigated by means of latex agglutination tests. The binding of fibronectin, collagen and lactoferrin to the same 55 strains was also tested by using 125I-labelled proteins. The 207 strains, belonging to ten Bacteroides species, were isolated from different infections (51%) and from faeces of healthy subjects (49%). Most of the strains displaying fibronectin binding belonged in the species B. fragilis or B. vulgatus. The binding could be inhibited by preincubation of the cells with an excess amount of fibronectin. No inhibition of the binding was observed with carbohydrates. The vitronectin binding of the strains was less common, but was always observed to accompany fibronectin binding. None of the examined 55 strains exhibited any binding to fetuin or asialofetuin. The radiolabelling method indicated a low binding to 125I-fibronectin. The binding of 125I-collagen-I and 125I-lactoferrin in the Bacteroides strains tested was higher than that of 125I-fibronectin.

Binding of extracellular matrix proteins to the surface of anaerobic bacteria.

The binding of fibronectin, vitronectin, collagen and sialoprotein to 65 anaerobic strains was investigated by means of latex agglutination tests. The binding of fibronectin, collagen and lactoferrin to the same strains was also tested by means of 125I-labelled proteins. The strains were isolated from abdominal infections (55%), from the faeces of healthy subjects (29%) or from the depths of tonsils removed at tonsillectomy (16%). The binding of fibronectin and collagen to Bacteroides fragilis strains, tested by the latex agglutination assay, was stronger than their binding to other species. The vitronectin binding of the strains was less common, but was always accompanied by fibronectin binding. Binding to fibronectin-coated beads was inhibited by pre-incubation of the bacterial cells with soluble fibronectin and by heat or protease treatment of the bacterial suspension. No inhibition of the binding was observed with carbohydrates. None of the 65 strains exhibited any binding to fetuin or asialofetuin; 8% of the strains had a binding site for mucin. The binding to mucin-coated beads was inhibited by pre-incubation of the cells with mucin. The radiolabelling method indicated a low binding to 125I-fibronectin. The binding of 125I-collagen-I and 125I-lactoferrin was higher for the anaerobic strains tested.

Long-term results of anterior levatorplasty for fecal incontinence. A retrospective study.

PURPOSE: To review the long-term results of anterior levatorplasty for fecal incontinence. METHODS: Fifty-four women with obstetric trauma and 31 with idiopathic incontinence responded to a questionnaire 1.5 to 18.5 (median, 8.5) years after anterior levatorplasty. Results were classified as excellent, good, fair, or poor. RESULTS: An excellent or good result was reported in 40 of 54 (74 percent) patients with an obstetric injury and in 14 of 31 (45 percent) patients in the idiopathic group (P < 0.01). The presence of a cloaca (P < 0.05) and a young age (P < 0.05) were associated with a favorable outcome in the obstetric and idiopathic group, respectively. Length of follow-up and preoperative severity of incontinence were not significantly related to outcome. CONCLUSIONS: This study suggests that every second patient undergoing anterior levatorplasty for fecal incontinence has a successful result that is sustained in the long term. Obstetric trauma, presence of a cloaca, and young age are associated with a successful outcome.

Expression of platelet-derived growth factor beta receptor in chondrogenesis of perichondrial transplants.

An experimental study was done in two rabbits to establish whether the early stages of cartilage generation involves stimulation via the platelet-derived growth factor beta receptor (PDGFR-beta). Rib perichondrium was dissected off and transplanted to the knee joint. The perichondrial grafts were removed for microscopic investigation at day 0 and day 6 and kept frozen until analysed by immunohistochemistry and in situ hybridisation for the presence of PDGF beta receptor protein and mRNA, respectively. Samples from day 0 showed sparse cells with a positive immunohistochemical reaction. In samples from day 6 there were signs of perichondrial proliferation and chondroid maturation and a more widespread immunohistochemical staining reaction could be seen mainly on proliferating perichondrial cells, but also on some chondrocytes. In situ hybridisation confirmed the expression of PDGF beta receptor mRNA in the same areas of serial sections. These findings may indicate that PDGF beta receptor activity is part of the early response after autotransplantation of perichondrium.

A comparison between netilmicin with metronidazole and doxycycline as prophylaxis in elective colorectal surgery.

The aim of this study was to compare the prophylactic effects of netilmicin plus metronidazole administered for one day and of doxycycline administered for five days on the incidence of postoperative sepsis in patients undergoing elective colorectal surgery. One hundred patients were studied, 50 men and 50 women, with a mean age of 66.6 years. The patients were randomized into the two groups. Wound infections occurred in a total of 9 patients, 5 in the doxycycline group and 4 in the group given netilmicin plus metronidazole, i.e. the overall infection rate was 9%. It is concluded that there is no statistically significant difference on the incidence of postoperative wound sepsis in patients undergoing elective colorectal surgery between the group given doxycycline for five days and the group given netilmicin plus metronidazole for one day.

Bacterial flora in semen before and after doxycycline treatment of infertile couples.

A group of men under investigation for infertility have been studied with regard to findings of aerobic and anaerobic bacteria in their ejaculates before and after medication. The group consisted of 27 men who received doxycycline concomitantly with their partners according to a fixed treatment schedule. Prior to medication 23 men delivered ejaculates which demonstrated bacterial growth. A total of 53 isolates were obtained. Following doxycycline treatment a significant number of men had a reduction in the number of isolates. The reduction was mainly confined to peptococci and propionibacteria. Of the 9 men whose partners subsequently conceived, 6 harboured peptococci and/or propionibacteria in their semen before but not after treatment. It is concluded that these results warrant more extensive studies of the possible part played by peptococci and propionibacteria in genital-tract infections in the male partners of infertile unions.

Preoperative cervical microbial flora and post-abortion infection.

With the aim to find criteria for the prediction of the patients who are at risk of developing a post-abortion infection the pretreatment cervical microbial flora was compared between one series of patients who developed and another series of patients who did not develop such an infection. Aerobic and anaerobic bacteria as well as mycoplasma and fungi, were studied in 104 patients. The distribution of aerobic and anaerobic bacteria was similar in the 14 patients who later developed post-abortal infection and in uncomplicated cases. It is concluded that the cultivation of the cervical microbial flora cannot serve as a basis for the prediction of which patients will develop subsequent genital infections.