A duplex qPCR for the simultaneous detection of Escherichia coli O157:H7 and Listeria monocytogenes using LNA probes.

UNLABELLED: In this study, a duplex qPCR assay was developed for the needs of the Irish fish industry to screen for the two major food-borne pathogens of fish, Listeria monocytogenes and Escherichia coli O157:H7. The assay can claim positive or negative results for two pathogens in one go in only 20 h including 16 h universal pre-enrichment and compared to traditional ISO approved plate culture methods the labour and the cost involved in testing of one sample is reduced to minimum. The highly specific genomic areas targeted for PCR amplification in the assay are the hly gene for listeriolysin O (LLO) of L. monocytogenes and the stx gene for Shiga-like toxin expressed by E. coli O157:H7. The detection limit of the assay is consistent with the consumer protection limits of 1 pg genomic DNA or 1 CFU 25 g(-1) fish meat (with enrichment) allowing the test to be considered as a substitute to standard plate culture methods. SIGNIFICANCE AND IMPACT OF THE STUDY: The study highlights a novel duplex qPCR for Listeria monocytogenes and Escherichia coli O157:H7 that could be used as an alternative to plate-based ISO or singleplex PCR methods while minimizing the costs. The assay uses rapid DNA extraction methods and locked nucleic acid probes. Sensitivity and specificity are 100 and 98·95% respectively. The potential for quantitative rage of the assay is 10(8) -10(1) CFU ml(-1) .

A young adult male with peripheral artery disease.

A 19-year-old man was admitted with severe symptoms suggestive for bilateral occlusive disease of his lower extremities. Clinical, laboratory, and radiological evaluation established the diagnosis of Buerger's disease. Buerger's disease often poses a diagnostic challenge and requires a high degree of suspicion. Although rare for this patient's young age, it is important that a correct diagnosis is being made, because the disease process can be stopped if the patient stops smoking.

Near-total completion gastrectomy for severe postvagotomy gastric stasis: analysis of early and long-term results in 62 patients.

The aim of this study was to evaluate results of completion gastrectomy for severe postgastrectomy gastric stasis. A total of 51 women and 11 men underwent completion gastrectomy for gastric stasis between 1985 and 1996; follow-up was complete in 98% at 5.4 +/- 5 years. All patients had modified Visick scores preoperatively of grade III (37%) or IV (63%). Presentation included combinations of nausea, vomiting, postprandial pain, chronic abdominal pain, and chronic narcotic use. All had undergone prior vagotomy and had a median of four previous gastric operations. Hospital mortality was zero. Complications occurred in 25 patients (40%) and included the following: narcotic withdrawal syndrome (18%), ileus (10%), wound infection (5%), intestinal obstruction (2%), and anastomotic leak (5%). All or most symptoms were relieved in 43% (Visick grade I or II), but 57% of the patients remained in Visick grade III or IV. Nausea, vomiting, and postprandial pain were reduced from 93% to 50%, 79% to 30%, and 58% to 30%, respectively (P<0.05), but chronic pain, diarrhea, and dumping syndrome were not significantly affected. Univariate analysis revealed no preoperative characteristic to be predictive of good outcome. Logistic regression analysis suggested that the combination of nausea, need for total parenteral nutrition, and retained food in the stomach predicted a poor outcome (P<0.05). Completion gastrectomy is successful in 43% of patients. The combination of nausea, need for total parenteral nutrition, and retained food at endoscopy are negative prognostic factors.

Management of the leaking rectal stump after Hartmann's procedure.

Hartmann's procedure is often selected in situations in which the construction of a colorectal anastomosis is considered unsafe. Paradoxically, the creation of the rectal stump involves placement of an intestinal suture line that is prone to leakage. We report our experience with 11 patients with a clinically significant leakage from the rectal stump after Hartmann's procedure, which was manifested as persistent postoperative intra-abdominal infection. A method for the treatment and prevention of this potentially lethal complication is presented.

Predictive factors of the outcome of traumatic cervical spine fracture in children.

The uncommon traumatic cervical spine fractures and dislocations were studied in 227 consecutively treated children (1 to 17 years of age). Lower cervical spine injuries (C-3 to C-7) affected 73% of our patients. However, among the 38 patients younger than 8 years of age, 87% had an injury at the C-3 level or higher (P < .0001). There were 19 fatalities (8.4%), all of which were associated with injuries at the C-4 level or higher. Of the 11 patients with atlanto-axial fracture or dislocation, all died soon after the injury. All had an unstable fracture and cord transection that resulted in cardiorespiratory collapse. Injuries at the vertebral levels C-1, C-2, C-3, and C-4 were associated with fatality rates of 17%, 9%, 4.3%, and 3.7%, respectively. No fatalities were encountered among patients with lesions lower than C-4. The authors conclude that the younger the age, the higher the cervical spine injury, and that the fatality rate correlates directly with the level of cervical spine fracture.

Pomegranate juice flavonoids inhibit low-density lipoprotein oxidation and cardiovascular diseases: studies in atherosclerotic mice and in humans.

The beneficial health effects attributed to the consumption of fruit and vegetables are related, at least in part, to their antioxidant activity. Of special interest is the inverse relationship between the intake of dietary nutrients rich in polyphenols and cardiovascular diseases. This effect is attributed to polyphenols' ability to inhibit low-density lipoprotein (LDL) oxidation, macrophage foam cell formation and atherosclerosis. Pomegranate polyphenols can protect LDL against cell-mediated oxidation via two pathways, including either direct interaction of the polyphenols with the lipoprotein and/or an indirect effect through accumulation of polyphenols in arterial macrophages. Pomegranate polyphenols were shown to reduce the capacity of macrophages to oxidatively modify LDL, due to their interaction with LDL to inhibit its oxidation by scavenging reactive oxygen species and reactive nitrogen species and also due to accumulation of polyphenols in arterial macrophages; hence, the inhibition of macrophage lipid peroxidation and the formation of lipid peroxide-rich macrophages. Furthermore, pomegranate polyphenols increase serum paraoxonase activity, resulting in the hydrolysis of lipid peroxides in oxidized lipoproteins and in atherosclerotic lesions. These antioxidative and antiatherogenic effects of pomegranate polyphenols were demonstrated in vitro, as well as in vivo in humans and in atherosclerotic apolipoprotein E deficient mice. Dietary supplementation of polyphenol-rich pomegranate juice to atherosclerotic mice significantly inhibited the development of atherosclerotic lesions and this may be attributed to the protection of LDL against oxidation.

[Posttraumatic superficial temporal artery pseudoaneurysm].

The etiology of superficial temporal artery aneurysm (STAA) is usually blunt trauma. This type of aneurysm is more prevalent in young men and is also probably more common than estimated from the literature. We present 3 males and 1 female with STAA, ranging in age from 8 to 30 years, who were successfully treated. 3 were operated on under local anesthesia, while the aneurysm of the fourth was obliterated by continuous local pressure. Application of local pressure is the best measure for preventing the development of pseudoaneurysm following blunt temporal trauma. We consider surgery the treatment of choice for STAA.

[Update of experience with prostatic cancer].

A retrospective analysis of 302 patients with prostatic cancer, referred between 1977-1982, was updated. 15 patients (5%) with Stage A1 disease did not receive any specific treatment, and their 10-year actuarial survival was 77%. 22 (7%) had Stage A2 disease at diagnosis, 82 (27%) Stage B, 88 (29%) Stage C, and 95 (32%) Stage D. Their 10-year actuarial survival rates were 75%, 56%, 38%, and 2%, respectively. The 10-year actuarial survival of patients with well- and moderately well-differentiated adenocarcinoma was 61%, but only 13% in those with poorly differentiated adenocarcinoma (p < 0.001). A total of 116 patients with Stages A2, B and C were treated by irradiation and 71 of them also received prophylactic hormone therapy. The 10-year actuarial survival of those treated by combined irradiation and hormone therapy was 59%, compared to 19% for those treated by irradiation alone (p < 0.02). Our results emphasize the need for early diagnosis of prostatic cancer, using modern diagnostic tools such as transrectal ultrasonography and determination of prostatic specific antigen. They also prove that radiotherapy is effective and curative in a high percentage of patients with disease confined to the pelvis.

[Rectus sheath hematoma].

Rupture of the epigastric artery with hematoma formation in the rectus sheath is an uncommon condition which can mimic serious intraabdominal disorders. Treatment is primarily conservative, but misdiagnosis can lead to unnecessary surgery. However, the correct clinical diagnosis should be suggested by the presence of paroxysmal cough, anticoagulant therapy, or trauma, the precipitating factors found in most cases. Ultrasonography and CT scan are excellent diagnostic modalities in this condition. 3 unoperated cases in which the correct diagnosis was made are presented. Despite its benign nature this condition may be fatal. Awareness and vigorous supportive therapy are therefore recommended to reduce morbidity and mortality.

[Trash foot following abdominal aortic surgery].

Peripheral embolism from dislodgement of intraarterial debris can convert successful surgery of the abdominal aorta into tragedy. An embolic shower from such surgery, often containing cholesterol, can result in forefoot or toe ischemia referred to as "trash foot." The incidence of this early complication is reported as 7-29%, but is believed to be even higher. Prevention and treatment includes heparin, embolectomy, lumbar sympathectomy, aspirin, rheomacrodex and prostaglandins. 3 such patients, 2 men aged 62 and 69, respectively, and a woman of 72, are reported. In 2 of the cases below-knee amputation was required, while the third patient died.

[Temporal artery biopsy--required or superfluous?].

Temporal arteritis is a systemic disease affecting large and medium-sized arteries in the elderly. The incidence of the disease increases with age and its major complications are blindness, cerebrovascular accidents and aortic dissection. Diagnosis is mainly based on clinical signs and symptoms. Temporal artery biopsy is a popular and simple diagnostic procedure and if positive confirms the diagnosis. However, a negative biopsy cannot exclude temporal arteritis due to its segmental nature, and the specific signs and symptoms still require treatment with corticosteroids. During the years 1982-1991 we performed 206 temporal artery biopsies, of which only 21 (10.2%) confirmed the presence of temporal arteritis. Our experience is presented with regard to the usefulness of temporal artery biopsy in particular. In view of the low biopsy yield we recommend more selective referral for this purpose.

[Subtotal cholecystectomy: an emergency procedure for the difficult gallbladder and high-risk patient].

Emergency cholecystectomy in high-risk patients is still associated with significant morbidity and mortality. Occasionally technical difficulties and bleeding diathesis are complicating factors. Our prospective experience with subtotal cholecystectomy in 23 consecutive patients is presented. All presented as increased surgical risks (APACHE II above 10) and suffered from acute cholecystitis with empyema or perforation. 1 patient died (4.4%), but overall, surgical complications were minimal. We conclude that subtotal cholecystectomy combines the advantages of cholecystectomy and cholecystostomy. We believe that this short, simple and safe procedure is a logical choice for emergency situations in critically ill patients.

[Isolated iliac aneurysm].

The incidence of isolated iliac artery aneurysm is 1-2% of that of abdominal aortic aneurysms. The natural history is of gradual enlargement, with rupture the most common clinical presentation. The signs and symptoms of such an aneurysm are influenced by its concealed location within the bony pelvis. Awareness of these special characteristics improves the chances of early diagnosis and proper surgical treatment before possible rupture. We report 2 cases which demonstrate the spectrum of the clinical presentation.

[Total colectomy and mucosal proctectomy with J-pouch anal anastomosis for ulcerative colitis and familial colonic polyposis].

Total colectomy and mucosal proctectomy with ileal pouch-anal anastomosis is the accepted surgical procedure for ulcerative colitis and familial polyposis of the colon. During 1981-1990, 25 patients with ulcerative colitis or familial polyposis underwent this operation in our department. In the majority a J-pouch was performed. In the early years, an 8 cm rectal muscular sleeve was left. In later cases, in accordance with opinions expressed in the medical literature, the length of the sleeve was shortened to about 3 cm. We present the functional results and the early and late complications on follow-up of up to 10 years (mean 3.5 years). Although this operation is not the ideal solution, it is better than the alternatives and is the surgical procedure of choice.