Sexual quality of life of adolescents and young adult breast cancer survivors.

BACKGROUND: With increasing survival rates of adolescents and young adults (AYAs) with breast cancer, health-related quality of life (HRQoL) becomes more important. An important aspect of HRQoL is sexual QoL. This study examined long-term sexual QoL of AYA breast cancer survivors, compared sexual QoL scores with that of other AYA cancer survivors, and identified factors associated with long-term sexual QoL of AYA breast cancer survivors. MATERIALS AND METHODS: Data of the SURVAYA study were utilized for secondary analyses. Sexual QoL was assessed using the European Organization for Research and Treatment of Cancer Quality of Life cancer survivorship core questionnaire (EORTC QLQ-SURV100). Descriptive statistics were used to describe sexual QoL of AYA cancer survivors. Linear regression models were constructed to examine the effect of cancer type on sexual QoL and to identify factors associated with sexual QoL. RESULTS: Of the 4010 AYA cancer survivors, 944 had breast cancer. Mean sexual QoL scores of AYA breast cancer survivors ranged from 34.5 to 60.0 for functional domains and from 25.2 to 41.5 for symptom-orientated domains. AYA breast cancer survivors reported significantly lower sexual QoL compared to AYA survivors of other cancer types on all domains. Age, time since diagnosis, relationship status, educational level, chemotherapy, hormonal therapy, breast surgery, body image, and coping were associated with sexual QoL of AYA breast cancer survivors. CONCLUSIONS: AYA breast cancer survivors experience decreased sexual QoL in the long term (5-20 years) after diagnosis and worse score compared to AYA survivors of other cancer types, indicating a clear need to invest in supportive care interventions for those at risk, to enhance sexual well-being.

Simultaneous use of an (125)I-seed to guide tumour excision and (99m)Tc-nanocolloid for sentinel node biopsy in non-palpable breast-conserving surgery.

PURPOSE: In the present study we describe patients with non-palpable breast lesions, in which an Iodine-125 ((125)I)-marker (or "seed") for excision of the primary tumour and Technetium-99m nanocolloid ((99m)Tc-nanocolloid) for sentinel node biopsy (SNB) are used simultaneously. The purpose was to investigate any interference between (125)I-seeds and (99m)Tc-nanocolloid by an in vitro and in vivo analysis. METHODS: Contrast/interference-ratios between (125)I and (99m)Tc count-rates were determined in vitro using a realistic simulation model. Measurements were performed with 3 gamma-probes with different crystal materials. In 25 consecutive patients (99m)Tc-nanocolloid was intratumourally administered at the site of a previously implanted (125)I-seed. Respectively, the (125)I-setting and (99m)Tc-setting of the gamma-probe guided the wide local excision and SNB and maximum counts-per-second (cps) were measured. RESULTS: In vitro the different probes varied in (125)I- and (99m)Tc-sensitivity. The contrast-ratio between (125)I and (99m)Tc in the (125)I-channel was 4.6 for a 3-month-old (125)I-seed using the most appropriate gamma-probe. In vivo the gamma-probe in the (125)I-setting measured a median of 16,300 cps at the tumour site compared to 4820 cps using the (99m)Tc-setting. The (125)I-seed could be well distinguished from the (99m)Tc-nanocolloid in 92% of the patients and 96% required a single operation. The SNB was successful in all patients. CONCLUSIONS: Simultaneous use of (125)I-seeds and (99m)Tc-nanocolloid is possible under well-standardised conditions. Non-palpable breast lesions can be safely excised using the (125)I-seed in combination with a SN procedure. Use of (125)I-seeds is a next step within fine-tuning breast-conserving surgery that should lead to further investigation to confirm its value.

Lymphoscintigraphy and SPECT/CT in multicentric and multifocal breast cancer: does each tumour have a separate drainage pattern? Results of a Dutch multicentre study (MULTISENT).

PURPOSE: To investigate whether lymphoscintigraphy and SPECT/CT after intralesional injection of radiopharmaceutical into each tumour separately in patients with multiple malignancies in one breast yields additional sentinel nodes compared to intralesional injection of the largest tumour only. METHODS: Patients were included prospectively at four centres in The Netherlands. Lymphatic flow was studied using planar lymphoscintigraphy and SPECT/CT until 4 h after administration of (99m)Tc-nanocolloid in the largest tumour. Subsequently, the smaller tumour(s) was injected intratumorally followed by the same imaging sequence. Sentinel nodes were intraoperatively localized using a gamma ray detection probe and vital blue dye. RESULTS: Included in the study were 50 patients. Additional lymphatic drainage was depicted after the second and/or third injection in 32 patients (64%). Comparison of planar images and SPECT/CT images after consecutive injections enabled visualization of the number and location of additional sentinel nodes (32 axillary, 11 internal mammary chain, 2 intramammary, and 1 interpectoral. A sentinel node contained metastases in 17 patients (34%). In five patients with a tumour-positive node in the axilla that was visualized after the first injection, an additional involved axillary node was found after the second injection. In two patients, isolated tumour cells were found in sentinel nodes that were only visualized after the second injection, whilst the sentinel nodes identified after the first injection were tumour-negative. CONCLUSION: Lymphoscintigraphy and SPECT/CT after consecutive intratumoral injections of tracer enable lymphatic mapping of each tumour separately in patients with multiple malignancies within one breast. The high incidence of additional sentinel nodes draining from tumours other than the largest one suggests that separate tumour-related tracer injections may be a more accurate approach to mapping and sampling of sentinel nodes in patients with multicentric or multifocal breast cancer.

Conjunctival provocation with airborne allergen in patients with atopic keratoconjunctivitis.

BACKGROUND: Atopic keratoconjunctivitis (AKC) is a chronic eye disease with periods of exacerbations. Many patients experience no obvious seasonal variation, although a majority of patients are allergic to common airborne allergens. OBJECTIVE: To investigate the allergic reaction, to conjunctival provocation with airborne allergens, in patients with AKC. METHODS: Eleven patients with AKC and birch and/or grass pollen allergy participated in the study, which was performed outside the pollen season. Five patients with seasonal allergic conjunctivitis (SAC) and five healthy subjects were included for validation purposes. The challenge was performed in one eye with the allergen, to which the patient was reactive, and with dilution buffer in the other eye. Signs and symptoms from both eyes were graded at baseline and at 10 min, 8 and 48 h after provocation. Tear fluid was collected from both eyes for cytokine analyses at baseline and at 8 and 48 h. RESULTS: A significant change in clinical symptoms and signs, (redness and chemosis) was evident 10 min after provocation compared with baseline (P = 0.005) and compared with the unprovoked eye (P = 0.005) in AKC subjects. These parameters were normalized after 8 and 48 h. A significant increase for IFN-γ (P = 0.021) and IL-6 (P = 0.015), and a near significant increase for IL-10 (P = 0.066) were seen in the tear fluid of the challenged eye at 48 h after provocation vs. baseline and vs. the control eye for IFN-γ (P = 0.005), IL-6 (P = 0.028) and IL-10 (P = 0.008) in AKC subjects. CONCLUSION AND CLINICAL RELEVANCE: In this single dose allergen provocation study, AKC patients responded with a typical IgE-mediated allergic reaction. An increase in cytokines at 48 h after the challenge was demonstrated and might, with further studies, give us a better understanding of the nature of inflammation in AKC.

Ponseti casting: a new soft option.

We have modified the Ponseti casting technique by using a below-knee Softcast instead of an above-knee plaster of Paris cast. Treatment was initiated as soon as possible after birth and the Pirani score was recorded at each visit. Following the manipulation techniques of Ponseti, a below-knee Softcast was applied directly over a stockinette for a snug fit and particular attention was paid to creating a deep groove above the heel to prevent slippage. If necessary, a percutaneous Achilles tenotomy was performed and casting continued until the child was fitted with Denis Browne abduction boots. Between April 2003 and May 2007 we treated 51 consecutive babies with 80 idiopathic club feet with a mean age at presentation of 4.5 weeks (4 days to 62 weeks). The initial mean Pirani score was 5.5 (3 to 6). It took a mean of 8.5 weeks (4 to 53) of weekly manipulation and casting to reach the stage of percutaneous Achilles tenotomy. A total of 20 feet (25%) did not require a tenotomy and for the 60 that did, the mean Pirani score at time of operation was 2.5 (0.5 to 3). Denis Browne boots were applied at a mean of 10 weeks (4 to 56) after presentation. The mean time from tenotomy to boots was 3.3 weeks (2 to 10). We experienced one case of cast-slippage during a period of non-attendance, which prolonged the casting process. One case of prolonged casting required repeated tenotomy, and three feet required repeated tenotomy and casting after relapsing while in Denis Browne boots. We believe the use of a below-knee Softcast in conjunction with Ponseti manipulation techniques shows promising initial results which are comparable to those using above-knee plaster of Paris casts.

Radio guided occult lesion localization (ROLL) for non-palpable invasive breast cancer.

BACKGROUND: Wire guided localization (WGL) for non-palpable breast cancer is technically difficult and patient unfriendly. Radio guided occult lesion localization (ROLL) takes advantage of the possibility to detect the tumor through the nuclear tracer that is injected directly into the tumor for the sentinel node procedure. METHODS: Forty patients with 41 invasive breast carcinomas were treated using ROLL. Patients received a dose of 120 Mbq 99mTc Nanocolloid intra-tumorally on the day of surgery or a dose of 370 Mbq 99mTc Nanocolloïd intra-tumorally the prior day. The sentinel node (SN) was located using patent blue and a gamma ray detection probe that was also employed to guide the tumor excision. RESULTS: In 31 patients (78%) the invasive tumor was adequately excised. In two cases (5%) a re-excision was required due to inadequately excised carcinoma in situ and in three patients (7.5%) both the invasive and the in situ tumor were inadequately excised. In 35 patients (88%) the SN was found and removed. CONCLUSIONS: The ROLL procedure seems to be an alternative to WGL in patients with non-palpable breast carcinoma. To determine the place of ROLL versus WGL in the treatment of non-palpable breast cancer, a randomized clinical trial is needed.

Axillary recurrence after a tumour-negative sentinel node biopsy in breast cancer patients: A systematic review and meta-analysis of the literature.

BACKGROUND: Sentinel node biopsy became the standard of care before consensus on the technique was reached and without randomized studies having shown a similar or decreased axillary recurrence rate. The purpose of this study was to evaluate studies reporting on patients with a negative sentinel node biopsy. METHODS: We performed a systematic review and meta-analysis of the literature for studies concerning clinically node-negative breast cancer patients with a tumour-negative sentinel node biopsy and no subsequent axillary node dissection. The axillary recurrence rate was determined, as well as the sensitivity of the sentinel node procedure and the differences in lymphatic mapping techniques. RESULTS: Forty-eight studies concerning 14 959 sentinel node-negative breast cancer patients followed for a median of 34 months were selected. Sixty-seven patients developed an axillary recurrence, resulting in a recurrence rate of 0.3%. The sensitivity of the sentinel node biopsy was 100%. Uni- and multivariable variable analyses showed that the lowest recurrence rates were reported in studies performed in cancer centres, in studies that described the use of (99m)Tc-sulphur colloid, and also when investigators used the superficial injection technique or evaluated the harvested sentinel nodes with haematoxylin-eosin and immunohistochemistry staining (p<0.01). CONCLUSIONS: In this systematic literature review, the axillary recurrence rate in sentinel node-negative patients is 0.3%, which is well within the desired range. The median sensitivity of the procedure appears to be as high as 100%. The recurrence rate is influenced by the differences in the lymphatic mapping technique.

'Radioguided occult lesion localisation' (ROLL) for non-palpable breast lesions: a review of the relevant literature.

BACKGROUND: 'Radioguided occult lesion localisation' (ROLL) is a possible alternative to the commonly used 'wire-guided localisation' (WGL) of non-palpable breast lesions. Intratumoural injection of a radiotracer identifies both the primary tumour and the sentinel lymph nodes for intraoperative gamma probe guided dissection. METHOD: A broad literature search was conducted, using the Pubmed, Embase and The Cochrane Library databases. Five clinically controlled trials and one randomised controlled trial, all of modest size, were selected which describe both ROLL and WGL. Four articles were found that describe the combined use of the ROLL technique with the sentinel node procedure. RESULTS: In the ROLL group between 69% and 84% of the lesions were radically excised, compared with 44-60% of the lesions in the WGL group. The studies that combined ROLL and the sentinel node procedure mentioned even higher percentages of radically excised specimens ranging from 90% to 95% and an identification rate of sentinel nodes up to 100%. One randomised controlled trial mentioned that the ROLL procedure had a faster localisation time, was easier to perform, was less painful and gave a better cosmetic result than WGL. CONCLUSION: ROLL seems a promising technique, which appears to be more radical than WGL. Localisation tends to be more accurate and faster, the excision procedure is more elegant and simple to perform, and the cosmetic result seems to be better. Further research is necessary to establish the exact role of ROLL in the management of non-palpable breast lesions.

Radiofrequency ablation for breast cancer: a review of the literature.

INTRODUCTION: Radiofrequency ablation (RFA) provides an effective technique for minimally invasive tissue destruction. A novel application is the use for treatment of small breast carcinoma. METHODS: A broad search was conducted in Pubmed, Embase and the Cochrane library. Results of the relevant articles were analysed. RESULTS: The analysed studies were all feasibility or pilot studies using different patient and tumour characteristics and ablation settings. Despite many methodological differences, high percentages of complete tumour ablation varying between 80% and 100% were reported. CONCLUSION: Radiofrequency ablation is a promising new tool for minimally invasive ablation of small carcinomas of the breast. A large randomized control study is required to assess the long-term advantages of RFA compared to the current breast conserving therapies.

Tacrolimus ointment vs steroid ointment for eyelid dermatitis in patients with atopic keratoconjunctivitis.

AIMS: The main objective of this explorative study was to evaluate if tacrolimus ointment could be safer than corticosteroid ointment, with special reference to the intraocular pressure in the treatment of eyelid eczema in patients with atopic keratoconjunctivitis (AKC). Secondary aims were to compare the effects of the treatments on eyelid eczema and their potential impact on ocular surface inflammation. METHODS: Tacrolimus 0.1% ointment and clobetasone butyrate 0.05% ointment were compared in a double-masked explorative crossover study. In total, 25 AKC patients were included. Each ointment was applied twice daily for 3 weeks, with 2 weeks of washout before, between, and after treatments. Efficacy was determined by eye examination and the patients' own symptom scoring. Cytology and cytokine measurements were performed on tear samples. Safety parameters were intraocular pressure, presence of bacteria and fungi, and the patients' reports of adverse events. The validity of the crossover design was explored with analysis of variance, and the effect of each medication was calculated with paired t-test and Wilcoxon paired test. RESULTS: A total of 20 patients completed the study. Both treatments were effective in reducing signs and symptoms of eyelid eczema, with a near superior benefit for tacrolimus in terms of eczema (total skin score) signs (P=0.05). No serious adverse events occurred and interestingly, intraocular pressure was not evidently affected by either treatment. CONCLUSION: Tacrolimus 0.1% ointment is a promising alternative therapy for eyelid eczema in AKC patients. Long-term studies are needed to further determine the value of tacrolimus in this patient group.

No apparent association between periocular and ocular microcolonization and the degree of inflammation in patients with atopic keratoconjunctivitis.

BACKGROUND: The cause of the chronic inflammation in atopic keratoconjunctivitis (AKC), the ocular manifestation of atopic eczema/dermatitis syndrome, is largely unknown. OBJECTIVE: To investigate the possibility that microorganisms may be important in the inflammatory activity in AKC. METHODS: Fifteen patients with AKC participated in the study. The presence of aerobic bacteria and fungi was related to the severity of clinical signs, the numbers of inflammatory cells in tears and conjunctival biopsies, and the concentration of various cytokines in tears. In addition, serological evidence for IgE sensitization to Staphylococcus aureus B antigen and Malassezia sympodialis antigen was investigated. Twelve healthy subjects were included for control purposes. RESULTS: The patients exhibited moderate clinical signs of AKC. No relation was found between the severity of AKC and the presence of microorganisms, despite the fact that S. aureus was frequently isolated. AKC patients showed significantly higher levels of IFN-gamma, TNF-alpha (tumour necrosis factor-alpha), IL-2, IL-4, IL-5 and IL-10 than controls. An association was found between conjunctival signs and the levels of all cytokines except IL-5. CONCLUSION: We found no evidence to suggest that periocular and ocular microcolonization are related to inflammatory parameters in AKC. However, confirmation of the present results in a longitudinal study with repeated clinical examinations and samplings in the same individual is required before the contribution of S. aureus to on-going inflammation in AKC can be dismissed.

The tryptase inhibitor APC-366 reduces the acute airway response to allergen in pigs sensitized to Ascaris suum.

BACKGROUND: Tryptase is a mast cell serine protease that is released during mast cell degranulation. It has been implicated as an important enzyme in the pathophysiology of asthma, but its role in this disease is not fully elucidated. OBJECTIVE: In this study, we investigated the effects of a tryptase inhibitor, APC-366, on the acute allergic airway reaction in specific pathogen-free pigs sensitized to the antigen Ascaris suum. METHODS: APC-366 (5 mg in 1 mL of water, each dose) was given as an aerosol to seven pigs two times (t); at t = - 60 min and t = - 15 min Control pigs received water. Ascaris antigen (in 2 mL saline) was nebulized to the airways over approximately 5 min at t = 0. All aerosols were generated with an ultrasonic nebulizer. RESULTS: The allergen challenge caused an acute reaction with a significant increase in airway resistance (R(aw)) in the control pigs from 3.3 +/- 0.6 cmH20/l/s to 10.2 +/- 2.3 cmH20/l/s, while in the APC-366-treated pigs, the R(aw) increased from 2.6 +/- 0.4 cmH20/l/s to 4.5 +/- 0.7 cmH20/l/s (P < 0.05 compared to controls). The dynamic lung compliance (C(dyn)) decreased significantly in the control pigs, but not in the APC-366-treated animals. The histamine concentration in urine in the control pigs was elevated immediately after allergen challenge, while this release was markedly reduced in the APC-366-treated pigs. CONCLUSION: The tryptase inhibitor APC-366 reduces the acute airway response to allergen significantly. There is also a reduced elevation in urine histamine concentration after challenge in the treated pigs, compared to controls. These results indicate that inhibition of mast cell tryptase might be a useful anti-allergic treatment in asthma.

Similar T helper Th2-like cytokine mRNA expression in vernal keratoconjunctivitis regardless of atopic constitution.

BACKGROUND: Many patients with vernal keratoconjunctivitis (VKC), a severe chronic allergic eye disease in children, exhibit IgE-sensitization, but about 40% of cases lack this immunologic trait. As a disease factor in VKC, IgE is thus not fully understood. The aim of this study was to investigate whether there are any differences in the conjunctival cytokine messenger (m)RNA pattern related to IgE-sensitization in children suffering from VKC. METHODS: Tissue samples were obtained from 16 symptomatic VKC subjects with sub-tarsal disease and six control subjects. Expression of mRNA for interleukin (IL)-4, IL-5, IL-13, and interferon (IFN)-gamma was investigated with a semiquantitative reverse-transcriptase polymerase chain reaction (RT-PCR) technique. The presence of T cells, IgE+ cells, mast cells, and eosinophils was analyzed with immunohistochemical methods. Allergen-specific IgE antibodies were assessed in serum and with skin prick testing. RESULTS: Ten out of the 16 VKC subjects showed evidence of IgE-sensitization. No differences were detected for any tissue variable between VKC subjects with and without IgE-sensitization. Statistically significant increases over controls were found for both VKC groups with regard to all cell markers. CONCLUSIONS: The amount of messenger RNA encoding cytokines and inflammatory cell markers in VKC did not correlate with IgE-sensitization. Tissue changes in all patient samples were characterized by a prevalence of T cells, eosinophils, mast cells and cell-bound IgE molecules. However, the role of cell-bound IgE molecules in VKC patients lacking IgE-sensitization remains to be determined.

IgE is expressed on, but not produced by, fetal cells in the human placenta irrespective of maternal atopy.

The prevalence of atopic diseases in children has increased during the last decades. Atopic symptoms usually appear early in life. This implies an early priming for atopic disease, possibly even at the fetal level. We therefore compared the presence and production of IgE in the local in utero environment during pregnancy in atopic and non-atopic women. Eighty-six women were included in the study. Fifty women were demonstrated to be atopics, based on clinical symptoms of atopic disease together with a positive Phadiatop and/or skin prick test. Placentas from these term pregnancies were obtained. Slices covering the full thickness of the placenta were cut clockwise around the umbilical cord and were analysed with immunohistochemistry. Surprisingly, numerous IgE+ cells, located primarily in the fetal villous stroma, were detected in a majority of the investigated placentas irrespective of the atopy of the mother or maternal or fetal total serum IgE levels. The placental IgE could not be demonstrated to be bound to IgE receptors, but was shown to be bound to fetal macrophages, possibly via FcgammaRI. No evidence was found for local fetal IgE production, although cells producing epsilon transcripts were occasionally detected in the decidua. We describe here the novel finding of numerous IgE+ cells in the human placenta, suggesting an hitherto unknown role for IgE in a successful pregnancy outcome, irrespective of whether or not the mother is atopic.

The effect of a bradykinin B2 receptor antagonist, NPC-567, on allergen-induced airway responses in a porcine model.

OBJECTIVE AND DESIGN: In order to assess the effect of selective blocking of the bradykinin (BK) B2 receptor in allergic airway reactions, the BK B2 receptor antagonist NPC-567 was administered to sensitized pigs before allergen challenge. MATERIAL: Fourteen specific pathogen-free pigs sensitized to Ascaris suum were used. TREATMENT: NPC-567 (2.5 mg, in 1 ml saline) was delivered as an aerosol twice to six pigs. METHODS: Ascaris antigen (in 2 ml saline) was given as an aerosol to all pigs and airway mechanics were monitored for 8 h. NPC-567 (2.5 mg) was given at t = -30 min (in 1ml saline) and mixed with the antigen at t = 0 to six pigs. RESULTS: Allergen challenge caused an acute reaction with a rapid, significant increase in airways resistance from 4.1 +/- 0.5 cm H2O/l/s to a maximum of 16.2 +/- 3.0 cm H2O/l/s in the control pigs. In the NPC-567-treated pigs, the resistance only increased from 2.9 +/- 0.3 cm H2O/l/s to 6.5 +/- 0.9 cm H2O/l/s (p<0.005 compared to controls). There was also a higher reduction in dynamic lung compliance in the controls than in the treated animals upon allergen challenge. The histamine concentration in urine in the control pigs was markedly elevated after allergen challenge peaking at 15-30 min. This release was inhibited in the NPC-567-treated pigs. CONCLUSIONS: The BK B2 receptor antagonist NPC-567 seems to be effective in inhibiting the acute response to allergen in the pig airways, possibly due to inhibition of mast cell activation via indirect mechanisms. The late obstructive response was reduced as well, probably as a consequence of the reduced mediator release in the acute reaction.

Inhibition of macrophage proinflammatory cytokine expression by steroids and recombinant IL-10.

Chronic lung disease (CLD) of prematurity is a prolonged respiratory failure in very-low-birth-weight neonates. Proinflammatory cytokines have been implicated in the development of CLD. Steroids have been shown to produce some improvement in neonates with this disease. The purpose of this study was to evaluate the downregulation of these proinflammatory cytokines by dexamethasone, budesonide and recombinant IL-10 (rIL-10) in order to elucidate the mechanism of the clinical benefit of steroids in babies. Our results showed that dexamethasone, budesonide and rIL-10 significantly inhibited both IL-6 and TNF-alpha production in the THP-1 cell line stimulated by lipopolysaccharide and Ureaplasma urealyticum antigen. Similar effects were found in macrophages from tracheobronchial aspirate fluid from newborn infants. In the rat alveolar macrophage cell line, steroids inhibited IL-6 and TNF-alpha production, while rat rIL-10 did not significantly decrease production. In conclusion, steroids and human rIL-10 were able to downregulate proinflammatory cytokine production, which may explain the beneficial effect of steroids and suggests that rIL-10 could be tried as an anti-inflammatory agent in neonates with a high risk of CLD.

Cytokine mRNA expression in patients with mild allergic asthma following low dose or cumulative dose allergen provocation.

BACKGROUND: Allergen provocation is a very useful way to study the inflammatory response in asthmatic patients. Although cumulative dose regimens are most often applied, another provocation model with repeated inhalations of low doses of allergens has recently come into use. OBJECTIVE: We were interested to compare these two allergen provocation models. To evaluate the inflammation induced by either model, we examined the mRNA expression of several cytokines that are implicated in the orchestration of the inflammatory response observed in asthma. METHODS: Interleukin (IL)-4, IL-5, IL-13 and interferon (IFN)-gamma mRNA expression was analysed in bronchoalveolar lavage (BAL) cells and peripheral blood (PB) CD4+ and PB CD8+ T cells following any of the two provocation regimens. IL-4 and IFN-gamma mRNA expression was analysed by a competitive reverse transcriptase-polymerase chain reaction (RT-PCR) method, while IL-5 and IL-13 were analysed semiquantitatively, before and after allergen provocation with either model. RESULTS: After low dose provocations none of the cell populations studied showed a clear change in the pattern of IL-4 or IFN-gamma gene expression. In contrast, after cumulative dose provocation there was a clear tendency towards an increased IL-4 mRNA expression in BAL cells, correlating with a significant increase in IL-4 mRNA in PB CD4+ as well as in CD8+ T cells (P = 0.005 and P = 0.04, respectively). Regardless of the allergen provocation method used, in PB IL-4 mRNA was preferentially expressed by CD4+ cells while IFN-gamma was expressed more by CD8+ cells. IL-5 transcripts increased after low dose provocations in PB CD4+ T cells in six of eight patients, while after cumulative dose provocation IL-5 mRNA increased in BAL cells in six out of nine patients but decreased especially in PB CD8+ T cells in six out of eleven patients, suggesting an accumulation of IL-5 expressing cells to the lungs. CONCLUSION: Thus, the cumulative dose provocation regimen can induce a more pronounced Th2-like immune response in asthmatic patients than the low dose provocation model.

Respiratory syncytial virus and chlamydia are not detectable by PCR in ongoing vernal keratoconjunctivitis.

Respiratory syncytial virus (RSV) and chlamydial infection may be pathogenetic factors in allergic diseases, perhaps also in ocular allergy. We analyzed the presence of RSV and chlamydial nucleic acids using reverse transcription (RT)-PCR and PCR, respectively, in conjunctival biopsies from patients with vernal keratoconjunctivitis (VKC) in order to determine whether these agents play a role in the maintenance of the disease. All biopsy samples were negative for both RSV (n = 15 for VKC and n = 10 for control subjects) and chlamydia (n = 8 for VKC and n = 7 for control subjects) homologous sequences. A direct association between RSV or chlamydial infection and ongoing inflammation in VKC could, therefore, not be confirmed.

Analysis of epsilon germline transcripts and IL-4 mRNA expression in the adenoids suggests local IgE switching.

BACKGROUND: We have previously observed more frequent occurrence of IgE+ and FcepsilonRI+ cells in adenoids of atopic than nonatopic children. To investigate the hypothesis that the adenoids are involved in IgE production, we analyzed the levels of epsilon germline (epsilonGL), IL-4, and IFN-gamma transcripts in the adenoids in relation to atopy and presence of ear disease. METHODS: Adenoidectomy was performed on 19 atopic and 18 nonatopic children (median age 5 years, range 2-12 years) suffering from otitis media with effusion (OME) (n = 16) or obstructive adenoids hyperplasia (AH) (n = 21). The levels of epsilonGL transcripts, IL-4, and IFN-gamma mRNA were analyzed by competitive reverse transcriptase-PCR. RESULTS: EpsilonGL transcript levels in the adenoids were found to be dependent on IL-4 mRNA expression (P < 0.01) and serum IgE levels (P < 0.05) (R2 = 0.32, n = 37). IL-4 mRNA expression was associated with epsilonGL transcript levels (rs = 0.32, P = 0.05, n = 37), especially among patients with AH (rs = 0.53, P = 0.01, n = 21). No significant differences in IL-4 and IFN-gamma mRNA levels were observed between the groups. CONCLUSIONS: This study supports an IL-4-induced class switch to IgE production in the adenoids that might be of importance for inflammatory reactions in the upper respiratory tract.