Predictive value of DNA repair gene expression for response to neoadjuvant chemotherapy in breast cancer

Genome-wide analysis using microarrays has revolutionized breast cancer (BC) research. A substantial body of evidence supports the clinical utility of the 21-gene assay (Oncotype DX) and 70-gene assay (MammaPrint) to predict BC recurrence and the magnitude of benefit from chemotherapy. However, there is currently no genetic tool able to predict chemosensitivity and chemoresistance to neoadjuvant chemotherapy (NACT) during BC treatment. In this study, we explored the predictive value of DNA repair gene expression in the neoadjuvant setting. We selected 98 patients with BC treated with NACT. We assessed DNA repair expression in 98 formalin-fixed, paraffin-embedded core biopsy fragments used at diagnosis and in 32 formalin-fixed, paraffin-embedded post-NACT residual tumors using quantitative reverse transcription-polymerase chain reaction. The following genes were selected: BRCA1, PALB2, RAD51C, BRCA2, ATM, FANCA, MSH2, XPA, ERCC1, PARP1, and SNM1. Of 98 patients, 33 (33.7%) achieved pathologic complete response (pCR). The DNA expression of 2 genes assessed in pre-NACT biopsies (PALB2 and ERCC1) was lower in pCR than in non-pCR patients (P=0.005 and P=0.009, respectively). There was no correlation between molecular subtype and expression of DNA repair genes. The genes BRCA2 (P=0.009), ATM (P=0.004), FANCA (P=0.001), and PARP1 (P=0.011) showed a lower expression in post-NACT residual tumor samples (n=32) than in pre-NACT biopsy samples (n=98). The expression of 2 genes (PALB2 and ERCC1) was lower in pCR patients. These alterations in DNA repair could be considered suitable targets for cancer therapy.

Atividades biológicas de espécies amazônicas de Rubiaceae / Biological activities of Amazonian species of Rubiaceae

RESUMORubiaceae é uma família botânica com grande potencial químico e biológico Neste trabalho realizou-se a prospecção fitoquímica e avaliou-se as atividades citotóxica, antioxidante e antifúngica dos extratos diclorometânicos e metanólicos das folhas de Duroia saccifera, Ferdinandusa goudotiana, F. hirsuta, F. paraensis, Ferdinandusa sp., Palicourea corymbifera e P. guianensis. A avaliação da citotoxicidade foi realizada empregando-se o ensaio de toxicidade sobre Artemia salina. A atividade antifúngica foi determinada pelo método de difusão em ágar utilizando-se os fungos: Epidermophyton floccosum, Microsporum canis 32905, M. gipseum 29/00, Trichophyton mentagrophytes ATCC 9533/03, T. rubrum ATCC 28189, T. tonsurans 21/97, Cladosporium cladosporioides, C. sphaerospermum, FusariumU. 662/06, Scytalidium U. 661/06, Candida albicans ATCC 3632 e C. albicans U. 5/99. A atividade antioxidante foi determinada através dos ensaios de redução do radical livre 1,1-difenil-2-picrilhidrazila (DPPH) e de descoloração do cátion radical ácido-6-sulfônico-2,2-azinobis-3-etilbenzotiazolina (ABTS). Para o ensaio de toxicidade sobre A. salina observou-se maior toxicidade no extrato metanólico de F. goudotiana, que se mostrou tóxico até a concentração de 5 µg mL-1. A presença de princípios antifúngicos foi observada em F. hirsuta e F. paraensiscontra quatro dos fungos testados. Na avaliação da atividade antioxidante os extratos metanólicos mostraram maior atividade que os extratos diclorometânicos, sendo coincidente o resultado obtido através de ambos os métodos (redução do DPPH e descoloração do ABTS). Os resultados criam possibilidades para futuras investigações relacionadas à estrutura e atividade dos componentes de cada extrato ativo.

ABSTRACTRubiaceae is a plant family with great chemical and biological potential. In this work a phytochemical prospection was performed and it was assessed the cytotoxic, antioxidant and antifungal activities of dichloromethanic and methanolic extracts of the leaves of Duroia saccifera, Ferdinandusagoudotiana, F. hirsuta, F. paraensis, Ferdinandusa sp., Palicourea corymbifera and P. guianensis. The cytotoxicity evaluation was carried out using the test toxicity on Artemia salina. The antifungal activity was determined by agar diffusion method using fungi: Epidermophyton floccosum, Microsporum canis 32905, M. gipseum 29/00, Trichophyton mentagrophytes ATCC 9533/03, T.rubrum ATCC 28189, T. tonsurans 21/97 , Cladosporium cladosporioides, C. sphaerospermum, FusariumU. 662/06, Scytalidium U. 661/06, Candida albicans ATCC 3632 and C. albicans U. 5/99. The antioxidant activity was determined by testing the amount of free radical 1,1-diphenyl-2-picrylhydrazyl (DPPH) and the discoloration of the radical cation 6-sulfonic acid-2 ,2-azinobis-3-ethylbenzothiazoline (ABTS). For the toxicity test on A. salina greater toxicity in the methanol extract of F. goudotiana was observed, which proved to be toxic up to a concentration of 5 µg mL-1. The presence of antifungal principles was observed in the F. hirsuta and F. paraensis against four tested fungi. In the evaluation of the antioxidant activity, the methanol extracts showed greater activity than the dichloromethanic ones, being similar the obtained result through both methods (reduction of DDPH and ABTS decolorization) . The results create possibilities for future researches related to the structure and activity of the components of each active extract.

Imbalanced expression of functional surface molecules in regulatory and effector T cells in systemic lupus erythematosus

Regulatory T (TREG) cells play an important role in maintaining immune tolerance and avoiding autoimmunity. We analyzed the expression of membrane molecules in TREG and effector T cells in systemic lupus erythematosus (SLE). TREG and effector T cells were analyzed for the expression of CTLA-4, PD1, CD28, CD95, GITR, HLA-DR, OX40, CD40L, and CD45RO in 26 patients with active disease, 31 with inactive disease, and 26 healthy controls. TREG cells were defined as CD25+/highCD127Ø/lowFoxP3+, and effector T cells were defined as CD25+CD127+FoxP3Ø. The ratio of TREG to effector T cells expressing GITR, PD1, HLA-DR, OX40, CD40L, and CD45RO was determined in the three groups. The frequency of TREG cells was similar in patients with SLE and controls. However, SLE patients had a decreased frequency of CTLA-4+TREG and CD28+TREG cells and an increased frequency of CD40L+TREG cells. There was a decrease in the TREG/effector-T ratio for GITR+, HLA-DR+, OX40+, and CD45RO+ cells, and an increased ratio of TREG/effector-T CD40L+ cells in patients with SLE. In addition, CD40L+TREG cell frequency correlated with the SLE disease activity index (P=0.0163). In conclusion, our findings showed several abnormalities in the expression of functionally critical surface molecules in TREG and effector T cells in SLE that may be relevant to the pathogenesis of this disease.

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.

Cytomegalovirus anterior uveitis: outcome of treatment.

AIM: To determine the outcome of antiviral treatment of cytomegalovirus (CMV) anterior uveitis. METHODS: A retrospective review of patients from Singapore National Eye Centre with CMV anterior uveitis diagnosed by aqueous polymerase chain reaction. Ganciclovir treatment consisted of systemic, topical, intravitreal injections or intravitreal implant. The main outcome measure was resolution of anterior chamber inflammation. RESULTS: 72 eyes of 70 patients were positive for CMV DNA. 35 eyes were treated (23 eyes with acute recurrent anterior uveitis and 12 eyes with chronic anterior uveitis). Eyes that did not respond or recurred with one treatment may receive another course of treatment. There were 47 treatment episodes, 36 (76.6%) of which resulted in a response. However, there were 27 (75.0%) episodes of recurrences after stopping treatment. Systemic and intravitreal ganciclovir and ganciclovir implant had good response rates but also had very high recurrence rates. Ganciclovir gel had moderate response rates, but its recurrence rates were also lower than those of the other modalities. CONCLUSIONS: Ganciclovir gel had lower recurrence rates than the systemic ganciclovir and the implant and should be considered as an option for treatment of CMV anterior uveitis.

Risk factors for endogenous Klebsiella endophthalmitis in patients with Klebsiella bacteraemia: a case-control study.

AIM: To identify risk factors for Klebsiella endophthalmitis in patients with Klebsiella sepsis. METHODS: This is a retrospective case-control study. The study population consisted of all patients with positive blood cultures for Klebsiella, admitted to Changi General Hospital (Singapore) from August 2004 to July 2005. The cases were patients who developed Klebsiella endophthalmitis, and the controls were those who did not. The potential risk factors analysed included age, sex, race and total white cell count at presentation, maximum temperature response, co-morbid pathologies, presence of severe infection, site of primary infection, antibiotic sensitivity of the bacterial strain, and systemic antibiotics treatment regimen. RESULTS: 133 consecutive patients with Klebsiella bacteraemia confirmed by blood culture were included. Five (3.8%) developed endophthalmitis. Klebsiella endophthalmitis was significantly associated with liver abscess as the primary site of infection (p<0.001) and disseminated intravascular coagulation (p = 0.010) on both univariate and multivariate analysis. Diabetes mellitus and other co-morbid pathologies, race and maximum temperature response were not risk factors for the occurrence of endophthalmitis. CONCLUSIONS: Liver abscess and disseminated intravascular coagulation are risk factors for endogenous endophthalmitis in patients with Klebsiella sepsis.

Correlation between peripapillary atrophy and corticosteroid therapy in patients with Vogt-Koyanagi-Harada disease.

AIMS: To determine the correlation between systemic corticosteroid therapy and the occurrence and size of peripapillary atrophy (PPA) in patients with Vogt-Koyanagi-Harada (VKH) disease. METHODS: All patients with VKH disease were retrospectively reviewed for their corticosteroid regimen. The extent of the PPA, if present, was measured using digitized imaging software, by two masked observers. Eyes with myopia greater than 6 dioptres or glaucoma were excluded. The patients were classified into three groups: early high (EH), late high (LH), and low dose (LD), according to the dose and timing of corticosteroids received during the acute phase of the disease. RESULTS: There were 40 eyes in the EH group, 25 eyes in the LH group, and 23 eyes in the LD group. Multivariate analysis showed that corticosteroid therapy was the main determinant of PPA occurrence. All the eyes in the LD group had PPA and eyes in the LH groups were 4.02 times (95% confidence interval 1.24-13.07) more likely to develop PPA than those in the EH group. The LD group also had larger PPA to disc ratios than the EH group. (Mean of 2.83 vs0.19, P<0.001). CONCLUSION: The development and extent of PPA in patients with VKH disease appear to be dependent on the dose and timing of systemic corticosteroids.

Thanatophoric dysplasia type II with encephalocele and aortic hypoplasia diagnosed in an anatomical specimen.

A hitherto unknown combination of congenital anomalies was found in an anatomical specimen of a female neonate. External examination and additional CT and MRI studies showed thanatophoric dysplasia type II with cloverleaf skull and concomitant parietal meningoencephalocele and hypoplasia of the descending aorta. The possibilities of causal correlations are discussed.

Endogenous endophthalmitis.

Endogenous endophthalmitis still presents a diagnostic and therapeutic challenge even with improvements in therapeutic modalities. The main prognostic factor is the virulence of the causative organism: once the organism enters the eye, it rapidly destroys ocular tissues. With the increasing ability to prolong the life of very ill patients, incidence of endogenous endophthalmitis is expected to increase. Ophthalmologists must be ever more vigilant, because prompt treatment offers the best chance of saving the eye.

Is Posner Schlossman syndrome benign?

PURPOSE: To determine the clinical course of patients with Posner Schlossman syndrome (PSS). DESIGN: Retrospective noncomparative case series. PARTICIPANTS: Fifty-three eyes of 50 patients. METHODS: The case notes of all patients with PSS seen at the Uveitis Clinic of Singapore National Eye Centre were reviewed for evidence of glaucoma damage and risk factors. MAIN OUTCOME MEASURES: Visual field and optic disc changes consistent with glaucoma. RESULTS: There were 28 men and 22 women, and their mean age at onset was 35 years. Fourteen eyes (26.4%) were diagnosed to have developed glaucoma as a result of repeated attacks of PSS. Patients with 10 years or more of PSS have a 2.8 times higher risk (95% confidence interval 1.19-6.52) of developing glaucoma compared with patients with less than 10 years duration of the disease. Nine eyes (17%) underwent glaucoma filtering surgery with antimetabolites. Their postoperative follow-up ranged from 15 to 50 months (mean, 37 months). Four eyes continued to have episodes of iritis after surgery, and one of these eyes had elevated intraocular pressure during the event. CONCLUSIONS: A significant number of patients with PSS have glaucoma develop over time, and they need to have their optic disc appearance and visual fields carefully monitored.

Capsular block syndrome: A case series.

Capsular block syndrome (CBS) or capsular bag distension syndrome has been described after cataract removal with in-the-bag placement of a posterior chamber intraocular lens in the presence of an anterior continuous curvilinear capsulorhexis. Features of CBS include shallowing of the anterior chamber and an unexpected myopic overrefraction; occasionally, there is a persistent uveitis. The patient may be mistakenly diagnosed with pupil block glaucoma or endophthalmitis. We report 9 cases of CBS and their initial diagnoses and management. In 1 case, the capsular bag distention and anterior chamber shallowing are illustrated by ultrasound biomicroscopy.

Conjunctival rotation autograft for pterygium. An alternative to conjunctival autografting.

OBJECTIVE: To determine the safety and efficacy of conjunctival rotation autografting (CRA) as an alternative to conventional conjunctival autograft after pterygium excision. DESIGN: Prospective noncomparative case series. PARTICIPANTS: Consecutive patients seen at the Pterygium Clinic of the Singapore National Eye Centre who were thought to be unsuitable for conventional conjunctival autografting underwent a modified surgical procedure, which the authors describe as CRA. There were 51 rotation autografts performed on 45 eyes of 43 patients. INTERVENTION: In this procedure, the underlying fibrovascular pterygium tissue was removed and the original epithelium (with minimal subepithelial tissue included) replaced over the bare sclera with a 180 degrees rotation. Surgeries were performed by one surgeon (DT) from April 1995 to May 1996. MAIN OUTCOME MEASURE: Pterygium recurrence and complications of CRA were measured. RESULTS: The mean follow-up time was 12 months (range, 2-22 months). There were 46 primary and 5 recurrent pterygia. The indications for CRA were combined cataract and pterygium surgery (39.2%), double pterygia (31.4%), the need to preserve the superior conjunctiva (21.6%), and superior conjunctival scarring (7.8%). There were two recurrences (4% recurrence rate), one occurring at 4 months and the other occurring at 7 months after surgery. No significant complications were encountered. However, 50% of the grafts remained mildly injected for more than 3 months, and some remained injected for up to 13 months after surgery (average of 4 months). Pigmentary changes were also noted in six grafts (12%). CONCLUSION: Conjunctival rotation autografting is a useful technique of conjunctival grafting in cases in which it is not possible or desirable to use the superior conjunctiva as a donor source.

Test-retest variability of blue-on-yellow perimetry is greater than white-on-white perimetry in normal subjects.

PURPOSE: To compare long-term fluctuation of blue-on-yellow automated perimetry with white-on-white automated perimetry in normal subjects. METHODS: White-on-white and blue-on-yellow automated perimetry were performed on a Humphrey Visual Field Analyzer and an Octopus perimeter, both modified for blue-on-yellow perimetry. The study sample consisted of 31 eyes of 31 normal subjects for the Humphrey perimeter and 33 eyes of 33 normal subjects for the Octopus perimeter. After one practice session, each subject completed four testing sessions over a period of 2 to 8 weeks, each separated by at least 1 day. Each testing session consisted of both white-on-white and blue-on-yellow perimetry performed on one eye; the order of the tests was alternated for successive sessions. Long-term fluctuation (expressed as statistical variance) was calculated for each test location. Intersubject variability (expressed as statistical variance) across all subjects was determined for each test location. RESULTS: On the Humphrey perimeter, the long term fluctuation for blue-on-yellow perimetry (4.07 +/- 3.07 dB2) was significantly greater than that for white-on-white perimetry (1.97 +/- 0.99 dB2; P < .001). Long-term fluctuation increased as a function of eccentricity for both blue-on-yellow and white-on-white perimetry. Short-term fluctuation was significantly greater for blue-on-yellow (0.46 +/- 0.25 dB) than that for white-on-white perimetry (0.29 +/- 0.19 dB; P < .02). Finally, the intersubject variability was significantly greater in blue-on-yellow (13.2 +/- 2.8 dB2) than it was in white-on-white perimetry (4.25 +/- 1.13 dB2; P < .001). Similar results were found with the Octopus perimeter. CONCLUSIONS: Long-term fluctuation and short-term fluctuation of blue-on-yellow perimetry are greater than those of white-on-white perimetry in normal subjects. The increased long-term fluctuation requires appropriate statistical approaches when evaluating serial change of blue-on-yellow perimetry.

Incidence of acute primary angle-closure glaucoma in Singapore. An island-wide survey.

OBJECTIVES: To determine the incidence of acute primary angle-closure glaucoma (APACG) in Singapore and to identify demographic and meteorological risk factors. DESIGN: A prospective, island-wide incidence study. SETTING: All government and private ophthalmological practices in Singapore, from March 1, 1995, to February 29, 1996. METHODS: New cases of APACG were identified by all ophthalmologists in Singapore during a 1-year period. Demographic and clinical details were recorded. RESULTS: One hundred eighty-nine people (208 eyes) were seen with APACG for the first time during the 1-year period. These new cases represent an incidence of 12.2 per 100,000 per year (95% confidence interval, 10.5-13.9) in those aged 30 years and older. Major risk factors identified were female sex (relative risk, 2.4), Chinese ethnic origin (relative risk, 2.8), and age of 60 years or older (relative risk, 9.1). Half of those affected were seen 3 days or more after the onset of symptoms. Attacks were more frequent on hotter days. There also was a relationship between the number of attacks per day and the mean number of sunspots and mean solar radio flux during the previous 30 days. CONCLUSIONS: There is a high incidence of APACG in Singapore, with elderly women being the highest risk group. Chinese Singaporeans are at higher risk than other ethnic groups (Malay and Indian). There is often a substantial delay before these patients consult a physician. The onset of APACG seems to be associated with meteorological factors.

Cataract surgery after trabeculectomy.

BACKGROUND AND OBJECTIVE: To determine the effect of cataract surgery on glaucomatous eyes with good intraocular pressure (IOP) control after trabeculectomy. PATIENTS AND METHODS: Twenty-two eyes with functional blebs that underwent cataract extraction were retrospectively analyzed. RESULTS: The mean (+/- SD) preoperative IOP was 11.0 +/- 4.3 mm Hg. The mean (+/- SD) postoperative IOPs at 1, 2, 6, and 9 months were 15.5 +/- 4.9, 12.6 +/- 4.7, 14.6 +/- 5.6, and 19.0 +/- 7.9 mm Hg, respectively. At each interval except for the second month, the mean IOP was statistically significantly higher than the preoperative value (P = .0003, .24, .02, and .0009, respectively). The total number of medications was also higher (3 preoperatively versus 27 postoperatively). The interval between the two surgeries had no influence on IOP control. Intraoperative complications during cataract surgery, particularly vitreous loss, were associated with poor IOP control. Phacoemulsification had less of an effect on the postoperative IOP control than did extracapsular cataract extraction. CONCLUSIONS: Eyes with previous successful trabeculectomies had higher IOPs and required more medications after subsequent cataract surgeries.

Mass screening for diabetic retinopathy--a report on diabetic retinal screening in primary care clinics in Singapore.

INTRODUCTION: Mass screening for diabetic retinopathy is expensive and inaccessible if done by institutional ophthalmalogists. Most diabetics are seen in primary care. Hence it is logical to provide mass screening in primary care clinics. In Singapore, government polyclinics are ideal centres of screening as they are well organised and accessible to the community. SCREENING METHOD: An effective mass screening strategy must provide wide coverage, be low cost and have the ability to assess diabetic eyes accurately and quickly. Non-mydriatic fundal photography was used as the screening method. Mass coverage was achieved by rotating two cameras around six government polyclinics. Cost was reduced by training existing staff and organising the programme to provide a high turnover of screenees. The photographs were read by ophthalmologists in a government-owned hospital. Patients that required referral were referred to specialist eye clinics. RESULTS: A total of 13,296 patients were screened or rescreened during a period of 2 years (25 months). 2,911 patients or 21.8% of the total screened were found to have diabetic retinopathy. About half of these (10.8%) had sight threatening retinopathy. The most common sight threatening retinopathy was maculopathy (8.0%). Twenty-two percent of cases screened were referred. These include referral for other ocular conditions detected during the screening. CONCLUSION: Non-mydriatic fundal photography has proven to be both accessible and effective in screening diabetic eyes in urban Singapore and can be recommended for mass screening of diabetic eyes in the community.

Contact transscleral cyclophotocoagulation for end stage glaucoma.

Contact transscleral cyclophotocoagulation is a form of cyclodestructive procedure, which is effective in the lowering of intraocular pressure in eyes with refractory glaucoma by destroying the aqueous humor-producing ciliary processes. Continuous wave Nd:YAG (Neodymium: Yttrium-Aluminium-Garnet) laser is used in the cyclodestruction, the laser energy being delivered to the target tissues transsclerally via a fibre-optic probe. We used the Lasag microruptor III Nd:YAG laser to treat 16 cases of refractory glaucoma to evaluate the effectiveness of laser in pain relief and intraocular pressure control. The mean preoperative intraocular pressure of 48.3 +/- 9.3 mm Hg decreased to 30.6 +/- 17.4 mm Hg (p = 0.0001) during an average follow-up of 294.2 +/- 94.2 days (range, 181-465 days). Retreatment was required in eight of 16 eyes (50%). Four eyes (27%) had decreased vision from perception of light to no perception of light. None of the 15 patients developed phthisis bulbi or other serious complications. We conclude that transscleral cyclophotocoagulation is an effective modality of therapy for refractory and end stage glaucoma. The lower than recommended total energy we used in the treatment protocol was associated with lower incidence of complications but also lower success rate and higher retreatment rate.