New biomarkers of post-settlement growth in the sea urchin Strongylocentrotus purpuratus.

Some sea urchins, including the purple sea urchin Strongylocentrotus purpuratus, have been successfully used in aquaculture, but their slow growth and late reproduction are challenging to overcome when developing efficient aquaculture production techniques. S. purpuratus develops via an indirect life history that is characterized by a drastic settlement process at the end of a larval period that lasts for several weeks. During this transition, the bilateral larva is transformed into a pentaradial juvenile, which will start feeding and growing in the benthic habitat. Due to predation and other ecological factors, settlement is typically associated with high mortality rates in juvenile populations. Additionally, juveniles require several days to develop a functional mouth and digestive system. During this perimetamorphic period, juveniles use up larval resources until they are capable to digest adult food. Mechanisms underlying the onset of juvenile feeding and metabolism have implications for the recruitment of natural populations as well as aquaculture and are relatively poorly understood in S. purpuratus. The insulin/insulin-like growth factor signalling (IIS)/Target of Rapamycin (TOR) pathway (IIS/TOR) is well conserved among animal phyla and regulates physiological and developmental functions, such as growth, reproduction, aging and nutritional status. We analyzed the expression of FoxO, TOR, and ILPs in post-settlement juveniles in conjunction with their early growth trajectories. We also tested how pre-settlement starvation affected post-settlement expression of IIS. We found that FoxO provides a useful molecular marker in early juveniles as its expression is strongly correlated with juvenile growth. We also found that pre-settlement starvation affects juvenile growth trajectories as well as IIS. Our findings provide preliminary insights into the mechanisms underlying post-settlement growth and metabolism in S. purpuratus. They also have important implications for sea urchin aquaculture, as they show that pre-settlement nutrient environment significantly affects both early growth trajectories and gene expression. This information can be used to develop new biomarkers for juvenile health in sea urchin population ecology and aquaculture aquaculture.

Prevalence and genotyping of Cryptosporidium spp. in farm animals in Egypt.

In this study, we examined the prevalence and molecular characteristics of Cryptosporidium in buffalo, dairy cattle and sheep in different farms at Kafr El Sheikh Province, Egypt. Rectal fecal samples, including 466 samples from buffalo, 1697 from cattle and 120 from sheep, were collected from different ages and screened by modified Ziehl-Neelsen acid-fast microscopy for detection of Cryptosporidium oocysts. All studied farms were positives with an overall prevalence of 1.29% in buffalo (4.17% in claves versus 0.48% in adults), 7.07% in cattle (6.90% in calves versus 10.20% and 6.10% in heifers and adults, respectively) and 2.50% in sheep (4.40% in lambs versus 1.30% in adults). PCR-RFLP analyses of small-subunit rRNA genes from positive specimens revealed the occurrence of C. parvum and C. ryanae in buffalo; C. parvum, C. ryanae, C. bovis and C. andersoni in cattle and only C. xiaoi in sheep. Genotypes distribution showed that C. ryanae was the dominant species (60.0%) followed by C. parvum (40.0%) in buffalo calves. Meanwhile, in cattle calves, C. parvum was the commonest species (74.23%) followed by C. ryanae (16.10%) and C. bovis (9.70%). Subtyping of C. parvum based on sequence analysis of the polymorphic 60 kDa glycoprotein gene locus showed the presence of subtypes IIdA20G1 and IIaA15G1R1 in both buffalo and cattle calves, addressing the potential role of calves in zoonotic cryptosporidiosis in Egypt.

Molecular profile of the antrochoanal polyp: up-regulation of basic fibroblast growth factor and transforming growth factor beta in maxillary sinus mucosa.

BACKGROUND: Various pathogenetic mechanisms have been proposed to explain the development of antrochoanal polyps (ACPs); however, the cause is still largely unknown. The aim of this study was to characterize the expression and the potential role of a battery of molecular markers in the development of ACPs. A prospective controlled study of a case series was performed. METHODS: Tissue samples of maxillary sinus mucosa were obtained from 14 patients with ACPs, 17 patients with chronic nonpolypoid maxillary sinusitis, and 4 patients with normal maxillary sinus mucosa; RNAs were extracted from the sinus mucosa, and semiquantitative reverse transcription-polymerase chain reaction was performed for basic fibroblast growth factor, transforming growth factor P, and mucin genes (MUC), MUC5AC, MUC5B, and MUC8, to investigate their expression. RESULTS: The expression of basic fibroblast growth factor and transforming growth factor beta was significantly higher in ACPs than in chronic rhinosinusitis and healthy mucosa. Meanwhile, the levels of expression of MUC genes were higher in ACPs and chronic rhinosinusitis compared with healthy mucosa. CONCLUSION: These findings suggest that ACPs may represent an inflammatory reaction caused by overproduction of tissue-derived growth factors in an inductive environment.

Up-regulation of MUC5AC and MUC5B mucin genes in nasopharyngeal respiratory mucosa and selective up-regulation of MUC5B in middle ear in pediatric otitis media with effusion.

OBJECTIVES: Mucins are the major component responsible for the gel-like properties of mucus secretion. Currently, little is known about the regulation of mucins in otitis media with effusion (OME). We investigated the expression of MUC5AC and MUC5B mucin gene messenger (m)RNAs in middle ear effusions as well as in tissue biopsies of the respiratory mucosa of the nasopharynx by means of reverse-transcription polymerase chain reaction (RT-PCR). STUDY DESIGN: A prospective, controlled, clinical trial. METHODS: The study group consisted of 25 children with a diagnosis of OME who were to undergo bilateral ventilation tube insertion with adenoidectomy. A sample of middle ear effusion (n = 50) as well as tissue biopsy from nasopharyngeal respiratory mucosa (n = 25) were obtained from each patient. For the nasopharyngeal mucosal biopsies, two control groups were made up from 30 children with healthy ears who underwent tonsillectomy +/- adenoidectomy operation; control A, 15 children with adenoid hypertrophy, and control B, 15 children with healthy nasopharynx. RNAs were extracted from effusion and tissue samples, and semiquantitative RT-PCR was performed for MUC5AC and MUC5B to investigate their expression. RESULTS: In the middle ear effusions, MUC5B mRNA expression was detected in 48 (96%), whereas MUC5AC mRNA was detected in 8 (16%). On semiquantitative analysis, MUC5AC/glyceraldehyde 3-phosphate dehydrogenase (GAPDH) and MUC5B/GAPDH mRNA ratios in the study group were significantly increased compared with those in both control groups (P < .05). CONCLUSIONS: The results suggest that the middle ear has a unique mucin gene expression pattern when compared with that of the nasopharynx. Up-regulation of MUC5B in the ear may play an important role in the pathogenesis of middle ear effusions.

Molecular assessment of neck dissections supports preserving level IIB lymph nodes in selective neck dissection for laryngeal squamous cell carcinoma with a clinically negative neck.

OBJECTIVES: It was the aim of this study to determine whether level IIB lymph nodes can be saved in elective neck dissection as a treatment for patients with squamous cell carcinoma (SCC) of the larynx. We present a prospective analysis of a case series. METHODS: Thirty-one patients with SCC of the larynx and without palpable lymph nodes at the neck who underwent an elective neck dissection were prospectively studied. The incidence of micrometastasis to level IIB lymph nodes after performing elective neck dissection was evaluated by nested reverse transcription polymerase chain reaction for cytokeratin (CK)19 and CK20 as well as by pathological examination. RESULTS: Nested reverse transcription polymerase chain reaction for CK19 and CK20 mRNA presented similar results but differed from the pathological examination. Of the 31 patients, 6 (19%) by pathological analysis and 9 (29%) by molecular analysis had lymph nodes positive for metastatic SCC. By molecular analysis, only 1 of the 31 patients had involvement of level IIB lymph nodes. CONCLUSIONS: This region may be preserved in elective neck dissection in patients with SCC of the larynx, so that accessory nerve dysfunction can be minimized and operative time can be saved.

Prevalence of Helicobacter pylori DNA in recurrent aphthous ulcerations in mucosa-associated lymphoid tissues of the pharynx.

OBJECTIVE: To determine the presence of Helicobacter pylori and, if detected, its potential prevalence in causing recurrent aphthous ulcers confined to mucosa-associated lymphoid tissues of the pharynx. DESIGN: Prospective, controlled clinical trial. SETTING: Otolaryngology Department of Tanta University Hospitals, Tanta, Egypt. PATIENTS: A total of 146 patients with recurrent multiple aphthous ulcers of the oral cavity and pharynx and 20 normal control subjects. INTERVENTIONS: Patients were assigned to group 1 (n = 58), in which the ulcers were strictly limited to the lymphoid tissues, or group 2 (n = 88), in which the ulcers were randomly distributed in the oral cavity and pharynx. Helicobacter pylori DNA was extracted from 3-mm-diameter tissue samples, and polymerase chain reaction amplifications were performed for the 16S ribosomal RNA gene. MAIN OUTCOME MEASURE: Positivity for H pylori. RESULTS: In group 1, 39 patients (67%) were positive for H pylori DNA, while in group 2, 9 patients (10%) were positive (chi(2) test, P<.001). It was not detected in any of the 20 control samples. CONCLUSION: Our results support a possible causative role for H pylori in recurrent aphthous ulcerations with a characteristic distribution and affinity to mucosa-associated lymphoid tissues of the pharynx.

Level IIb lymph nodes metastasis in elective supraomohyoid neck dissection for oral cavity squamous cell carcinoma: a molecular-based study.

OBJECTIVES: To identify the incidence of level IIb lymph nodes metastasis in elective supraomohyoid neck dissection (SOHND) as a treatment for patients with squamous cell carcinoma (SCC) of the oral cavity. STUDY DESIGN: Prospective analysis of a case series. METHODS: Forty-eight patients with SCC of the oral cavity and with no palpable lymph nodes at the neck who underwent an elective SOHND were prospectively studied. The incidence of micrometastasis to level IIb lymph nodes after performing elective SOHND was evaluated by pathologic examination and molecular analysis. RESULTS: Of the 48 patients, 15 (31%) by pathologic analysis and 22 (46%) by molecular analysis had lymph nodes positive for metastatic SCC. By molecular analysis, 5 (10%) of the 48 patients had involvement of level IIb lymph nodes. All patients with metastasis to level IIb lymph nodes have their primary lesions in the tongue and constituted 22% of patients with tongue lesions. There was no instance of isolated metastasis to level IIb lymph nodes without involvement of other nodes in the SOHND specimens. CONCLUSIONS: In this study, level IIb lymph node metastasis was only found in association with tongue carcinoma. Although this region may be preserved in elective SOHND in patients with SCC of the oral cavity, it should be included whenever the tongue is the primary site.