Oligonucleotide microarray-based detection and genotyping of Plum pox virus

Plum pox virus (PPV) is the most damaging viral pathogen of stone fruits. The detection and identification of its strains are therefore of critical importance to plant quarantine and certification programs. Existing methods to screen strains of PPV suffer from significant limitations such as the simultaneous detection and genotyping of several strains of PPV in samples infected with different isolates of the virus. A genomic strategy for PPV screening based on the viral nucleotide sequence was developed to enable the detection and genotyping of the virus from infected plant tissue or biological samples. The basis of this approach is a long 70-mer oligonucleotide DNA microarray capable of simultaneously detecting and genotyping PPV strains. Several 70-mer oligonucleotide probes were specific for the detection and genotyping of individual PPV isolates to their strains. Other probes were specific for the detection and identification of two or three PPV strains. One probe (universal), derived from the genome highly conserved 3' non-translated region, detected all individual strains of PPV. This universal PPV probe, combined with probes specific for each known strain, could be used for new PPV strain discovery. Finally, indirect fluorescent labeling of cDNA with cyanine after cDNA synthesis enhanced the sensitivity of the virus detection without the use of the PCR amplification step. The PPV microarray detected and identified efficiently the PPV strains in PPV-infected peach, apricot and Nicotiana benthamiana leaves. This PPV detection method is versatile, and enables the simultaneous detection of plant pathogens.     

Molecular characteristics of extended-spectrum beta-lactamases among gram-negative isolates collected in Cairo University Hospital

Phenotyping is commonly used for detection of extended-spectrum beta-lactamase (ESBL) production in gram-negative isolates. ESBLs are mainly coded for by four important genes, namely bla (TEM), bla (SHV), bla (CTX-M), and bla (OXA). Our aim in this study is to assess use of a multiplex PCR as a rapid method to identify four common genes responsible for ESBL production in different gram-negative isolates. All 793 clinical isolates are subjected to both screen and confirmatory testing for ESBL production using double disc synergy testing (DDST). Two hundred isolates with the ESBL phenotype are subjected to multiplex PCR for detection of the four genes bla (TEM, SHV, CTX-M, and OXA). The isolates were obtained from various clinical specimens: 68 (34 %) were isolated from urine cultures, 43 (21.5 %) from sputum, 26 (13 %) from wounds, 34 (17 %) from blood culture, 20 (10 %) from stool of healthy carrier and nine (4.5 %) from bronchoalveolar lavages. In this study, 83 isolates (41.5 %) were from outpatients (urine and stool specimens only), and the remaining 117 isolates (58.5) were from inpatients. By PCR technique, 181 isolates were found to be ESBL producers. blaTEM was the commonest genotype (39.2 %), followed by blaSHV (32.5 %) and blaCTX-M (30.9 %), either alone or in combination. Acinetobacter baumannii isolate had none of the ESBL genes. Eighteen (9.9 %) out of 181 isolates carried more than one type of beta-lactamase genes. Our study demonstrated rapid detection of bla (TEM, SHV, CTX-M, and OXA) in isolates belonging to Enterobacteriaceae and other nonfermenting clinical isolates using multiplex PCR. This genotypic method provided a rapid and efficient differentiation of ESBLs in the laboratory.     

HAART and anti-Koch’s impair sexual competence,sperm quality and offspring quality when used singly and in combination in male Wistar rats

This study investigated the impact of the administration of HAART and anti-Koch's, singly and in combination, on sexual competence and birth statistics. Adult male Wistar rats were randomised into distilled water-treated control, HAART-treated, anti-Koch's-treated and HAART + anti-Koch's-treated groups. The 56-day oral treatment led to impaired sexual competence evident by significantly reduced motivation to mate, prolonged latencies of mount, intromissions, ejaculations and post-ejaculatory interval, as well as reduced frequencies of mount, intromissions and ejaculations. This was accompanied by significant reductions in penile erection reflex and penile grooming. HAART and anti-Koch's, when administered singly or in combination, also led to significant reductions in the circulatory follicle-stimulating hormone, luteinizing hormone, testosterone and intratesticular testosterone, but a significant rise in prolactin. Also, HAART and/or anti-Koch's significantly reduced sperm count, sperm motility, sperm viability and spermatozoa with normal morphology. Furthermore, HAART and anti-Koch's, separately or in combination, significantly lowered fertility capacity, litter size and litter weight and offspring survival. The deleterious effects of these drugs were more pronounced when combined. Findings of the present study revealed that HAART and/or anti-Koch's impair sexual competence via a testosterone-dependent hyperprolactinemia-mediated mechanism. These events are associated with reduced fertility capacity, poor sperm quality and lowered offspring survival.     

Pregnancy-related admissions to the intensive care unit

We conducted a retrospective review of obstetric patients admitted to the intensive care unit at Al-Ain hospital during period January 1(st) 1997 to December 31(st) 2002, in order to identify the indications for admission and the outcome. A total of 60 patients were admitted during the six years. The frequency of admission was 2.6 per 1000 deliveries and obstetric patients represented 2.4% of all ICU admissions. Admission was planned in 11 patients (18%) and unplanned in 49 (82%). The mean (+/-SD) duration of stay in ICU was 1.6+/-1.5 days. The leading indications for admission were haemorrhage (28.4%) and preeclampsia/eclampsia (25%). Of the 60 admissions, 47 (78.4%) followed surgery. The mean APACHE II score was 5.0+/-3.0. Twenty-two patients (37%) had blood transfusions, and only two (3.3%) required ventilation. Of the 60 patients only 28 (46.7%) were deemed to have severe illness necessitating intensive care; the remaining 32 patients were suitable for high dependency care. The mean APACHE II score and duration of stay were significantly higher in these patients. There were two deaths, representing 3.3% of obstetric intensive care unit admissions. Our findings highlight the need for establishing a high dependency unit to avoid unnecessary admission to the intensive care unit and to ensure proper management.     

Catheter-direct thrombolysis versus pharmacomechanical thrombectomy for treatment of symptomatic lower extremity deep venous thrombosis

BACKGROUND: Rheolytic mechanical thrombectomy using the AngioJet catheter (Possis Medical, Minneapolis, MN) has been shown to be effective in the treatment of deep venous thrombosis (DVT). Additional infusion of thrombolytic agents via the device creates a novel treatment strategy of pharmacomechanical thrombectomy (PMT), which further enhances thrombectomy efficacy. The purpose of the current study was to compare the treatment outcome in patients with symptomatic DVT who underwent either catheter-directed thrombolysis (CDT) or PMT intervention. METHODS: During a recent 8-year period, clinical records of all patients with symptomatic lower leg DVT undergoing catheter-directed interventions were evaluated. Patients were divided into 2 treatment groups: CDT or PMT. Comparisons were made with regards to the treatment outcome between the 2 groups. RESULTS: A total of 93 patients who underwent 98 catheter-directed interventions for DVT were included in the study. Among them, CDT or PMT was performed in 46 (47%) and 52 (53%) procedures, respectively. In the CDT group, complete or partial thrombus removal was accomplished in 32 (70%) and 14 (30%) cases, respectively. In the PMT cohort, complete or partial thrombus removal was accomplished in 39 (75%) and 13 (25%) cases, respectively. Venous balloon angioplasty and/or stenting in the CDT or PMT groups was necessary in 36 (78%) and 43 (82%), respectively (difference not significant [NS]). Patients in the CDT groups underwent a mean of 2.5 venograms during the hospital course, in contrast to 0.4 venograms per patient in PMT cohorts (P < .001). Immediate (<24 hours) improvement in clinical symptoms in CDT and PMT groups was achieved in 33 (72%) and 42 (81%) cases, respectively (NS). Significant reductions in the intensive care unit (ICU) and hospital lengths of stay was noted in the PMT group (0.6 and 4.6 days) when compared to the CDT group (2.4 and 8.4 days). During follow-up visits, the primary patency rates at 1 year of CDT and PMT groups were 64% and 68%, respectively (NS). Hospital cost analysis showed significant cost reduction in the PMT group compared to the CDT group (P < .01). CONCLUSIONS: PMT with adjunctive thrombolytic therapy is an effective treatment modality in patients with significant DVT. When compared to CDT, this treatment provides similar treatment success with reduced ICU, total hospital length of stay, and hospital costs.     

Circulating Vascular Growth Factor (VEGF) Angiopoietin-1 (Angi-1) and Soluble Tie-2 Receptor in Pregnancy Complicated with Pre-eclampsia: A Prospective Study

Background

Preeclampsia is a leading cause of maternal and fetal/neonatal mortality and morbidity worldwide. Although the etiology of preeclampsia (PE) is still unclear, recent studies suggest that its major phenotypes, high blood pressure and proteinuria, are due in part to the disturbed angiogenic process.

Study Design

This study included the following groups: (1) women with normal pregnancies (n = 150), (2) patients with PE (n = 88), and (3) patients who delivered small growth for date (SGA) neonate (n = 50). Maternal serum concentrations of VEGF, Angi-1, and sTie-2 were measured by a sensitive immunoassay. Non-parametric statistics were used for analysis.

Results

The median maternal serum concentration of sVEGF and sAngi-1 was lower in normal pregnant women as compared to that in PE and SGA and the differences were statistically significant (P < 0.01). In contrast, there is a significant reduction in sTie-2 levels in PE and SGA groups as compared to that in normal pregnancy group (P < 0.01). Serum VEGF and Angi-1 were significantly higher in the late onset PE subgroup as compared to that in the early onset PE (P < 0.01), but sTie-2 was not significantly different in the 2 subgroups (P > 0.05). Serum VEGF, sAngi-1, and sTie-2 were significantly higher and Tie-2 was significantly lower in the severe PE subgroup as compared to that of the milder PE subgroup (P < 0.01 for all).

Conclusion

Patients with PE and those with SGA fetuses have lower median serum concentrations of sTie-2 and higher sVEGF and sAngi-1 than women with normal pregnancies. These findings lend support to the hypothesis that circulating angiogenic proteins may have an important biologic role in PE.     

Invasive IPMN and MCN: Same Organ, Sifferent Outcomes?

Background

The efficacy of surgery for invasive mucinous neoplasms is unclear. We examined the natural history of invasive mucinous cystic neoplasms (MCN) and invasive intraductal papillary mucinous neoplasms (IPMN) in patients who underwent pancreatic resection.

Methods

The Surveillance, Epidemiology, and End Results (SEER) database (1996–2006) was queried for cases of resected invasive MCN and IPMN. Demographics, tumor characteristics, and overall survival were examined using log-rank analysis and multivariate Cox regression model.

Results

Of 185 MCN cases and 641 IPMN cases, 73% and 48%, respectively, were women (P < 0.0001). Most (73%) IPMN were in the head of the pancreas; most (64%) MCN were in the tail/body (P < 0.0001). Lymph node metastasis was more common for IPMN than MCN (46% vs. 24%, P < 0.0001). Overall survival after resection was better for patients with stage I MCN vs. stage I IPMN (P = 0.0005), and it was better for patients with node-negative MCN vs. node-negative IPMN (P = 0.0061). There was no significant difference in survival of patients with stage IIA MCN vs. stage IIA IPMN (P = 0.5964), stage IIB MCN vs. stage IIB IPMN (P = 0.2262), or node-positive MCN vs. node-positive IPMN (P = 0.2263). Age older than 65 years (hazards ratio (HR) 1.71, P = 0.0046), high tumor grade (HR 2.68, P < 0.0001), higher T stage (HR 2.11, P < 0.0001), and IPMN histology (HR 1.90, P = 0.0040) predicted worse outcome in node-negative patients.

Conclusions

Our findings suggest that survival is better after resection of invasive MCN versus invasive IPMN when disease is localized within the pancreas, but this difference disappears in the presence of nodal metastasis or extrapancreatic extension.     

Treatment of Mammary Duct Fistula by Fistulectomy and Saucerization

This study was designed to assess the efficacy and long-term outcome of fistulectomy and saucerization for treatment of mammary duct fistulae. Mammary fistula is a chronic condition that represents the final step in what has been termed “mammary duct associated inflammatory disease sequence.” The treatment is primarily surgical and may include healing by secondary intention or primary closure with or without antibiotics. Reported series are small and often include variable surgical strategies applied without consistency. A consecutive series of 53 patients who had 59 mammary duct fistulae were treated by fistulectomy with saucerization. The median age was 32 years. Wounds were allowed to heal by secondary intention and antibiotics were not used. We reviewed the case records to establish the incidence of recurrent fistula and the time to complete healing. The long-term cosmetic outcome was determined by a postal survey. After a median follow-up of 6 years there had been no relapse in 92%. There was significant delay in healing in six cases (range: 10 to 30 weeks). Thirty-eight patients (83%) gave a definite history of regularly smoking between 10 and 20 cigarettes a day. Two thirds of the patients were either pleased or satisfied with the final cosmetic result of the surgery, but more than 90% said that it left them with some distortion of the nipple. Fistulectomy and saucerization achieves long-term cure in the majority of patients with mammary duct fistula, but it results in some degree of distortion of the nipple. The strong relationship between smoking and the occurrence of mammary duct fistulae is again demonstrated.