Inhibiting mutant KRAS G12D gene expression using novel peptide nucleic acid-based antisense: A potential new drug candidate for pancreatic cancer

KRAS mutations, which are the main cause of the pathogenesis of lethal pancreatic adenocarcinomas, impair the functioning of the GTPase subunit, thus rendering it constitutively active and signaling intracellular pathways that end with cell transformation. In the present study, the AsPC-1 cell line, which has a G12D-mutated KRAS gene sequence, was utilized as a cellular model to test peptide nucleic acid-based antisense technology. The use of peptide nucleic acids (PNAs) that are built to exhibit improved hybridization specificity and have an affinity for complementary RNA and DNA sequences, as well as a simple chemical structure and high biological stability that affords resistance to nucleases and proteases, enabled targeting of the KRAS-mutated gene to inhibit its expression at the translation level. Because PNA-based antisense molecules should be capable of binding to KRAS mRNA sequences, PNAs were utilized to target the mRNA of the mutated KRAS gene, a strategy that could lead to the development of a novel drug for pancreatic cancer. Moreover, it was demonstrated that introducing new PNA to cells inhibited the growth of cancer cells and induced apoptotic death and, notably, that it can inhibit G12D-mutated KRAS gene expression, as demonstrated by RT-PCR and western blotting. Altogether, these data strongly suggest that the use of PNA-based antisense agents is an attractive therapeutic approach to treating KRAS-driven cancers and may lead to the development of novel drugs that target the expression of other mutated genes.     

Tinea capitis in Benghazi, Libya

BACKGROUND: Tinea capitis is a worldwide problem. It affects mainly school age children. Late detection and improper treatment of the inflammatory type of this disease may result in disfigurement and permanent alopecia. The objectives of this study were to determine the prevalence, clinical types, and causative species of tinea capitis in Benghazi, Libya. METHODS: One hundred and ninety six patients with tinea capitis were enrolled in this study. Hair stumps and scales were collected from every case and exposed to direct microscopic examination using 10% potassium hydroxide solution, and cultivation on modified Sabouraud's dextrose agar with cyclohexamide and chloramphenicol. RESULTS: Tinea capitis accounts for 45% of all superficial fungal infection and 92% occurred in children below the age of 10 years. The gray patch type was the most common clinical variety (53.6%), followed by black dots, seborrhoid type, and kerion (25.5%, 10.2%, and 8.2%), respectively. Four patients with a clinical picture of alopecia areata-like lesion and one patient with a favus-type lesion were seen. Species identification revealed that Trichophyton violaceum was the most common causative agent, responsible for 49.4% of infection, followed by Microsporum canis (38.6%) and T. verrucosum (7.8%). From seven patients the isolate was a mixture of both T. violaceum and M. canis. CONCLUSIONS: There has been a dramatic decrease in the incidence of favus with complete disappearance of T. schoenleinii. T. verrucosum as a causative agent of tinea capitis in this area has been reported for the first time in this study.     

'Occlusal sketch'; a reliable technique for technicians to check that the occlusion of models is correct?

The aim of this study was to determine whether the occlusal sketch technique can be used as a reliable means of verifying the occlusal contacts on articulated casts. Occlusal markings were made on 30 sets of upper and lower dentate casts and these were recorded using the 'occlusal sketch' technique. Fifteen out of the total 30 pairs of casts were deliberately corrupted by the clinician so that they no longer matched the record on the 'occlusal sketch'. Three dental technicians were asked to judge in which of the 30 casts the occlusal contacts differed from those represented by the occlusal sketch. The results showed almost perfect agreement between the technicians in correctly identifying which casts had marked occlusal contacts which differed from those recorded on the occlusal sketch. The occlusal sketch provides a reliable way of communicating occlusal contacts that occur in a case, between clinician and technician.     

Stabilization splint therapy for the treatment of temporomandibular myofascial pain: a systematic review

The aim of this review is to establish the effectiveness of stabilization splint (SS) therapy in reducing symptoms in patients with myofascial pain. Searching of electronic databases, handsearching of relevant key journals, and screening of reference lists of included studies were undertaken. There was no language restriction, and unpublished research was sought. The selection criteria were randomized controlled trials comparing splint therapy to either no treatment or another active treatment. Data extraction and validity assessment were carried out independently and in duplicate. Studies were grouped according to treatment type. Twenty potentially relevant Randomized Controlled Trials (RCTs) were identified. Only twelve met the inclusion criteria. There is insufficient evidence either for or against the use of stabilization splint therapy over other active interventions for the treatment of temporomandibular myofascial pain. However, it appears that stabilization splint therapy may be beneficial for reducing pain severity at rest and on palpation and depression when compared to no treatment. The authors suggested the need for well conducted RCTs that pay attention to method of allocation, blind outcome assessment, sample size, and duration of follow-up. Various measures were adopted to assess the outcomes of treatment. Standardization of the methods used to measure outcomes of the treatment of myofascial pain should be established in future RCTs.     

Rupture of middle cerebral artery aneurysm in a neonate: case report and review of the literature

Intracranial hemorrhage because of rupture of a cerebral aneurysm is extremely rare in the neonatal period. Delayed diagnosis contributes to high mortality and morbidity.The authors report an extremely rare case of a middle cerebral artery aneurysm diagnosed and treated shortly after birth. Extensive review of the literature is presented. The patient died 4 years after surgery.     

Acquired jejuno-ileal diverticular disease: a diagnostic and management challenge

Background: Acquired jejuno‐ileal diverticular disease (JID), a result of abnormalities in the smooth muscle or myenteric plexus of the small bowel, is less rare than was once believed. Approximately 1.3% of the population has JID, of whom approximately 10% present with life‐threatening complications such as inflammation, perforation, bleeding, obstruction and malabsorption. Jejuno‐ileal diverticular disease can be diagnostically and therapeutically challenging, and complications are often diagnosed only at laparotomy, while the best management is not agreed on in the literature. To increase the awareness of this condition and its complications, the Auckland Hospital's experience of JID was reviewed. Methods: Retrospective review was carried out of the audit data and the discharge coding records of Auckland Public Hospital for the 5 year period leading to November 2001. Results: Nine cases with a variety of presentations were found. Those cases are described and a literature review of JID is provided. Conclusion: Jejuno‐ileal diverticular disease should be included in the differential diagnosis when dealing with surgical emergencies in the elderly presenting with features of bowel perforation, obstruction or bleeding.     

Congo/Crimean haemorrhagic fever in Iraq

Congo/Crimean haemorrhagic fever was recognized for the first time in Iraq in 1979. The first case was reported on 3 September 1979 and since then a further 9 patients have been investigated. Eight patients gave a history of previous contact with sheep or cattle, while 2 patients, a resident doctor and an auxiliary nurse, acquired their infections in hospital by direct contact with patients. The causal virus was isolated from patients'' blood and postmortem liver specimens. The virus isolates were found to be closely related if not identical serologically to members of the Congo/Crimean haemorrhagic fever virus group. Eight of the patients had no epidemiological relationship to one another and lived in widely separated areas around Baghdad and Ramadi (110 km to the west of Baghdad).     

A quantitative technique for growing human adult skeletal muscle in culture starting from mononucleated cells

A quantitative and reproducible technique for establishing primary surface cultures from normal and diseased human muscle is described. Successful cultures were prepared from both fresh muscle and that stored up to 96 hr at 4 °C. The CPK activity of the muscle cells ranged between 0.5–3.0 μmoles creatine per min per mg protein at 30 °C, thus indicating a high degree of differentiation. Spontaneous contractions were observed in 4 out of the 22 cultures established. Nerve cells were not required to achieve this level of differentiated function.No gross differences in plating efficiency, rate of myotube formation or CPK specific activity were found for the diseased muscle cells cultured so far. However, a 5–10-fold higher cell yield was obtained from muscles of patients with an inflammatory myopathy.The advantages of this technique for carrying out comparative studies on normal and dystrophic muscle cells are discussed.     

Contractile actions of proteinase-activated receptor-derived polypeptides in guinea-pig gastric and lung parenchymal strips: evidence for distinct receptor systems

1. We have measured the contractile activities and relative potencies (EC(50)s) of six thrombin PAR(1) receptor-derived receptor-activating peptides (PAR-APs): AparafluroFRChaCit-y-NH(2) (Cit-NH(2)); SFLLRNP(P7); SFLLRNP-NH(2) (P7-NH(2)); SFLLR (P5); SFLLR-NH(2) (P5-NH(2)); TFLLR-NH(2) (TF-NH(2)) and a PAR(2) receptor activating peptide [SLIGRL-NH(2) (SL-NH(2))] (a) in a guinea-pig lung peripheral parenchymal strip preparation and (b) in a gastric longitudinal smooth muscle preparation. 2. The relative potencies of the PAR-APs in the lung preparation (Cit-NH(2) congruent with TF-NH(2) congruent with P5-NH(2) > P7 congruent with P5 congruent with P7-NH(2); SL-NH(2) not active) differed appreciably from their relative potencies in the gastric preparation: Cit-NH(2) congruent with TF-NH(2) congruent with P7-NH(2) congruent with P5-NH(2) > P7 congruent with SL-NH(2). 3. The contractile actions of the PAR(1)-selective peptide, TF-NH(2) in the gastric preparation were entirely dependent on extracellular calcium and were blocked by tyrosine kinase inhibitors (genistein, tyrphostin 47/AG213, PP1) and by the cyclooxygenase inhibitor, indomethacin, whereas in the lung preparation, the PAR(1)-mediated contractile response was only partially dependent on extracellular calcium and was refractory to the actions of either tyrosine kinase inhibitors or indomethacin. 4. Partial sequencing of the PAR cDNAs detected by RT - PCR both in whole lung and in the peripheral parenchymal strip bioassay tissue demonstrated the presence of both PAR(1) and PAR(2) mRNA; the expression of PAR(2) was detected by immunohistochemistry. 5. The data point to the presence of distinct receptor systems for the PAR(1)-APs in guinea-pig lung parenchymal and gastric smooth muscle and indicate that PAR(2) does not regulate contractile activity in peripheral parenchymal guinea-pig lung tissue     

Ruptured thoracoabdominal aneurysm in a 27-year-old with hyper IgE syndrome

Hyperimmunoglobulin E syndrome is a rare primary immunodeficiency disorder defined by high serum immunoglobulin E titers and associated with characteristic infectious, dermatologic, skeletal, and dental abnormalities. We present the case of a 27-year-old man presenting acutely with a ruptured type IV thoracoabdominal aortic aneurysm. He was successfully treated by open operative repair and discharged with long-term prophylactic antibiotics. To our knowledge, this is the first such case reported in the English literature.