Investigation of the Lethal Effect of Purified Arginine Deiminase Purified from Lactobacillus plantarum p5 on Murine Mammary Adenocarcinoma and Vero cell Lines.

Cancer is one of the most serious diseases facing humanity; accordingly, it is urgent to find a cure that is rarely harmful to the patient as much as possible. It has been approved that arginine deiminase (ADI) can hydrolyze the plasma arginine to citrulline. This hydrolysis activity and reduction in the amount of intercellular arginine suppress lipopolysaccharide-induced nitric oxide synthesis. On the other hand, arginine depletion arrests the cell cycle at the G1 phase; therefore, ADI has been considered a powerful anticancer agent. The current study aimed to investigate the lethal effects of ADI purified from the Lactobacillus plantarum p5 strain on murine mammary adenocarcinoma and Vero cell lines. Anti-proliferative activity of ADI against murine mammary adenocarcinoma) AMN3) cell line was evaluated after different incubation times (3, 6, 24, 48, and 72 h) of exposure to 1 µg/mL of ADI, compared to Vero (non-cancer cell line) transformed cell line with same conditions. The autophagy process in cancer cells was recognized after three hours of incubation with ADI which was clearly observed in the AMN3 cell line under an inverted microscope. The first stages of the programmed cell death (apoptosis) pathway were only observed in AMN3 cells after 24 h of incubation with ADI, and this process continued with the time until they reached the last stages of apoptosis after 72 h of incubation. The results of the current study showed that the AMN3 cell line was auxotrophic for arginine because it could not produce it in the presence of enzyme which had a robust activity to kill these cancer cells; however, Vero non-cancer cell line survived in the presence of ADI because it had the ability to produce arginine.

Velopharyngeal insufficiency in patients without a cleft palate: important considerations for the ENT surgeon.

BACKGROUND: Velopharyngeal insufficiency is the inability to close the velopharyngeal port during speech and swallowing, leading to hypernasal speech and food regurgitation. OBJECTIVE: This study aimed to explore the aetiological factors contributing to the development of velopharyngeal insufficiency in a non-cleft paediatric population, especially following adenoidectomy. METHODS: A retrospective case review was conducted of all children without a known cleft palate, born between 2000 and 2013, who were referred to a tertiary cleft centre with possible velopharyngeal insufficiency. RESULTS: The data for 139 children diagnosed with velopharyngeal insufficiency following referral to the cleft centre were analysed. Thirteen patients developed the condition following adenoidectomy; only 3 of these 13 had a contributing aetiological factor. CONCLUSION: Velopharyngeal insufficiency is a rare but significant complication of adenoidectomy. The majority of patients who developed velopharyngeal insufficiency following adenoidectomy did not have an identifiable predisposing factor. This has important implications for the consent process and when planning adenoidectomy.

Phenotypic conservation in patients with X-linked retinitis pigmentosa caused by RPGR mutations.

IMPORTANCE: For patients with X-linked retinitis pigmentosa and clinicians alike, phenotypic variability can be challenging because it complicates counseling regarding patients' likely visual prognosis. OBJECTIVE: To evaluate the clinical findings from patients with X-linked retinitis pigmentosa with 13 distinct RPGR mutations and assess for phenotypic concordance or variability. DESIGN: Retrospective medical record review of data collected from 1985 to 2011. SETTING: Kellogg Eye Center, University of Michigan. PATIENTS: A total of 42 patients with X-linked retinitis pigmentosa with mutations in RPGR. Age at first visit ranged from 4 to 53 years, with follow-up ranging from 1 to 11 visits (median follow-up time, 5.5 years; range, 1.4-32.7 years, for 23 patients with >1 visit). MAIN OUTCOMES AND MEASURES: Clinical data assessed for concordance included visual acuity (VA), Goldmann visual fields (GVFs), and full-field electroretinography (ERG). Electroretinography phenotype (cone-rod vs rod-cone dysfunction) was defined by the extent of photopic vs scotopic abnormality. Qualitative GVF phenotype was determined by the GVF pattern, where central or peripheral loss suggested cone or rod dysfunction, respectively. Goldmann visual fields were also quantified and compared among patients. RESULTS: Each mutation was detected in 2 or more related or unrelated patients. Five mutations in 11 patients displayed strong concordance of VA, while 4 mutations in 16 patients revealed moderate concordance of VA. A definitive cone-rod or rod-cone ERG pattern consistent among patients was found in 6 of 13 mutations (46.2%); the remaining mutations were characterized by patients demonstrating both phenotypes or who had limited data or nonrecordable ERG values. Concordant GVF phenotypes (7 rod-cone pattern vs 4 cone-rod pattern) were seen in 11 of 13 mutations (84.6%). All 6 mutations displaying a constant ERG pattern within the mutation group revealed a GVF phenotype consistent with the ERG findings. CONCLUSIONS AND RELEVANCE: While VA and ERG phenotypes are concordant in only some patients carrying identical mutations, assessment of GVF phenotypes revealed stronger phenotypic conservation. Phenotypic concordance is important for establishing proper counseling of patients diagnosed as having X-linked retinitis pigmentosa, as well as for establishing accurate patient selection and efficacy monitoring in therapeutic trials.

HLA DRB1*130101-DQB1*060101 haplotype is associated with acute chest syndrome in sickle cell anemia patients.

We investigated the association of human leukocyte antigens (HLA) class II alleles and haplotypes with the pathogenesis of acute chest syndrome (ACS) in 186 sickle cell anemia (SCA) patients, of whom 58 had documented ACS (new pulmonary infiltrate, fever, and other associated clinical events) and 128 with a negative history of ACS, serving as controls. HLA DRB1* and -DQB1* genotyping was performed by polymerase chain reaction-sequence-specific priming. Of the DRB1* and DQB1* alleles analyzed, only DRB1*130101 (Pc < 0.001) was positively associated with ACS. DRB1*130101-DQB1*060101 haplotype was more prevalent among ACS patients (P = 0.018), thus conferring disease susceptibility. Specific HLA alleles and haplotypes may influence ACS risk in SCA patients, and specific HLA genotypes may be useful markers for identifying high-risk SCA ACS patients.

Evidence for HLA class II susceptible and protective haplotypes for osteomyelitis in pediatric patients with sickle cell anemia.

We investigated the association of human leukocyte antigen (HLA) class II alleles and haplotypes with the pathogenesis of sickle cell anemia (SCA) osteomyelitis. SCA patients comprised 42 patients with osteomyelitis and 150 patients without osteomyelitis; HLA-DRB1* and HLA-DQB1* genotyping was performed by polymerase chain reaction-sequence-specific priming (SSP). DRB1*100101 (P value corrected for the number of different alleles tested, Pc=0.003) was positively associated with osteomyelitis. At the haplotype level, DRB1*100101-DQB1*050101 (Pc=0.001) was more prevalent among patients, while DRB1*030101-DQB1*0201 (Pc=0.020) and DRB1*040101-DQB1*0302 (Pc=0.039) were more prevalent among SCA controls, thereby conferring disease susceptibility or protection to these haplotypes, respectively. These results show that specific HLA haplotypes influence SCA osteomyelitis risk and that specific HLA types may serve as markers for identifying SCA patients at high risk for osteomyelitis.

Alternative drugs against Trichomonas vaginalis.

To investigate the effect of drugs other than metronidazole, 3 non-pregnant women infected with Trichomonas vaginalis were treated with doxycycline, 2 x 200 mg/day for 1 week. Another 3 women were treated with praziquantel, single dose, 40 mg/kg body weight. No therapeutic effect was detected for either drug. In vitro, oxytetracycline led to death of T. vaginalis at a concentration of 15 mg in 0.5 mL medium. Extract of Myrtus communis caused death of T. vaginalis at pH 4.65, but failed to do so at pH 6.00. Extract of Eucalyptus comaldensis (50 mg in 0.1 mL medium) at pH 5.35 caused death of T. vaginalis after 24 hours.

Alternative drugs against Trichomonas vaginalis

To investigate the effect of drugs other than metronidazole, 3 non- pregnant women infected with Trichomonas vaginalis were treated with doxycycline, 2x200 mg/ day for 1 week. Another 3 women were treated with praziquantel, single dose, 40 mg/ kg body weight. No therapeutic effect was detected for either drug. In vitro, oxytetracycline led to death of T. vaginalis at a concentration of 15 mg in 0.5 mL medium. Extract of Myrtus communis caused death of T. vaginalis at pH 4.65, but failed to do so at pH 6.00. Extract of Eucalyptus comaldensis [50 mg in 0.1 mL medium] at pH 5.35 caused death of T. vaginalis after 24 hours

Preclinical efficacy and safety of pascolizumab (SB 240683): a humanized anti-interleukin-4 antibody with therapeutic potential in asthma.

The type 2 helper T cell (T(H)2) cytokine interleukin (IL)-4 is thought to play a central role in the early stages of asthma. In an effort to develop an antibody treatment for asthma that neutralizes the effects of IL-4, a murine monoclonal antibody, 3B9, was generated with specificity for human IL-4. In vitro studies demonstrated that 3B9 inhibited IL-4-dependent events including IL-5 synthesis, (T(H)2) cell activation and up-regulation of immunoglobulin E expression. 3B9 was then humanized (pascolizumab, SB 240683) to reduce immunogenicity in humans. SB 240683 demonstrated species specificity for both monkey and human IL-4 with no reactivity to mouse, rat, cow, goat or horse IL-4. Pascolizumab inhibited the response of human and monkey T cells to monkey IL-4 and effectively neutralized IL-4 bioactivity when tested against several IL-4-responsive human cell lines. Affinity studies demonstrated rapid IL-4 binding by pascolizumab with a slow dissociation rate. In vivo pharmacokinetic and chronic safety testing in cynomolgus monkeys demonstrated that pascolizumab was well tolerated, and no adverse clinical responses occurred after up to 9 months of treatment. Three monkeys developed an anti-idiotypic response that resulted in rapid pascolizumab clearance. However, in the chronic dosing study the antibody response was transient and not associated with clinical events. In conclusion, pascolizumab is a humanized anti-IL-4 monoclonal antibody that can inhibit upstream and downstream events associated with asthma, including (T(H)2) cell activation and immunoglobulin E production. Clinical trials are under way to test the clinical efficacy of pascolizumab for asthma.

Cryptosporidiosis among animal handlers and their livestock in Basrah, Iraq.

OBJECTIVE: To investigate the prevalence of cryptosporidiosis among groups at risk (animal handlers) and among domestic animals. DESIGN: Comparative study with zoonotic aspect. METHOD: Stool samples were collected from 60 animal handlers, 175 non-animal handlers and 198 domestic animals (60 cows, 45 sheep, 45 goats, 25 horses and 23 camels). Direct smear method and then formalin-ether sedimentation method were carried out for stool samples to detect intestinal parasites. Faecal smears were prepared from the sediment and stained by the modified Ziehl-Neelsen method for the recovery of red-pink oocysts of Cryptosporidium. RESULTS: Out of the 60 animal handlers, 30 (50%) were found to be positive for intestinal parasites compared to 26 (14.8%) of non-animal handlers (P < 0.01). Cryptosporidium oocysts were found to be excreted by three (5%) animal handlers and two (1.14%) of the non-animal handlers (P > 0.05). Cryptosporidiosis was also diagnosed in 20%, 13.3%, 17.7% and 12% of cattle, sheep, goats and horses respectively. No single positive case was detected among the examined camels. CONCLUSION: Veterinarians, butchers and breeders should be aware of the disease among farm animals in order to avoid great losses and to prevent its transmission to humans.

Intestinal parasites, including Cryptosporidium species, in Iraqi patients with sickle-cell anaemia.

Stool samples were obtained from individuals admitted to three hospitals in Basra during November 1997-May 1998. Of 40 patients with sickle-cell anaemia, 25 (62.5%) had parasitic infections. In the apparently healthy comparison group, 26 of 175 individuals (14.8%) had intestinal parasitic infections, a statistically significant difference. The most common intestinal parasites isolated in the sickle-cell patients were Blastocystis hominis (36%) and Giardia lamblia (28%). The isolation rate of Cryptosporidium species in sickle-cell patients (5%) was not significantly different from that in apparently healthy individuals (1.14%). We report for the first time the isolation of Isospora belli from a sickle-cell patient in Iraq and the Mediterranean region.

Intestinal parasites, including Cryptosporidium species, in Iraqi patients with sickle-cell anaemia

Stool samples were obtained from individuals admitted to three hospitals in Basra during November 1997-May 1998. Of 40 patients with sickle-cell anaemia, 25 [62.5%] had parasitic infections. In the apparently healthy comparison group, 26 of 175 individuals [14.8%] had intestinal parasitic infections, a statistically significant difference. The most common intestinal parasites isolated in the sickle-cell patients were Blastocystis hominis [36%] and Giardia lamblia [28%]. The isolation rate of Cryptosporidium species in sickle-cell patients [5%] was not significantly different from that in apparently healthy individuals [1.14%]. We report for the first time the isolation of Isospora belli from a sickle-cell patient in Iraq and the Mediterranean region

Risk factors for vaginal trichomoniasis among women in Basra, Iraq.

We investigated Trichomonas vaginalis infection among 352 women with vaginal discharge, 46 were found to be infected, an infection rate of 13%. There were no significant differences in the isolation rate of T. vaginalis in women according to occupation, educational level, economic status, age, marital status, parity, menstrual status and contraception use. The difference in the isolation rates of T. vaginalis in women with a history of abortion (7.6%) and in women with no history of abortion (15.7%) was statistically significant.

Risk factors for vaginal trichomoniasis among women in Basra, Iraq

We investigated Trichomonas vaginalis infection among 352 women with vaginal discharge, 46 were found to be infected, an infection rate of 13%. There were no significant differences in the isolation rate of T. vaginalis in women according to occupation, educational level, economic status, age, marital status, parity, menstrual status and contraception use. The difference in the isolation rates of T. vaginalis in women with a history of abortion [7.6%] and in women with no history of abortion [15.7%] was statistically significant

Characterization of the in vitro and in vivo activity of monoclonal antibodies to human IL-18.

IL-18 is a cytokine with potent IFN-gamma inducing activities as well as an important mediator of Th1 polarized immune responses. In this study we demonstrated that IL-18 induces the concentration-dependent production of the proinflammatory mediators IFN-gamma, IL-6, and GM-CSF, but not the anti-inflammatory cytokine, IL-10 from peripheral blood lymphocytes in the presence of mitogen. Three neutralizing IL-18 monoclonal antibodies (MAbs) were investigated, one of which (2C10) inhibited IL-18 bioactivity with an IC50 of 0.1 nM and had a K(D) of 3.9 x 10(-11) M. A NOD/SCID mouse model engrafted with human peripheral blood lymphocytes was developed to test the in vivo efficacy of this MAb. The IFN-gamma production induced by LPS administration was inhibited approximately 90% by prior dosing of MAb 2C10. The therapeutic utility of a high-affinity IL-18 MAb may be of benefit in Th1-driven autoimmune diseases such as rheumatoid arthritis and Crohn's Disease, where elevated levels of IL-18 have been observed.

Inhibition of CD23 processing correlates with inhibition of IL-4-stimulated IgE production in human PBL and hu-PBL-reconstituted SCID mice.

BACKGROUND: CD23, the low affinity serum immunoglobulin E (IgE) receptor, is upregulated on B cells following interleukin (IL)-4 stimulation and is concomitantly cleaved to generate soluble CD23 (sCD23) fragments with cytokine-like activity. OBJECTIVE: Compounds that selectively inhibit the proteolytic release of CD23 to generate sCD23 were assessed for their ability to inhibit IgE production in order to evaluate the contribution of sCD23 in the production of human IgE as well as the ability of such compounds to block IgE production. METHODS: IgE production was measured in IL-4-stimulated human peripheral blood lymphocytes (PBL) and PBL-reconstituted SCID mice in the presence of a broad-spectrum matrix metalloprotease (MMP) inhibitor, a compound selective for inhibition of CD23 processing over MMPs and an anti-CD23 mAb, MHM6. RESULTS: The two compounds were equipotent in inhibiting IgE production without inhibition of IgG production by IL-4/anti-CD40-stimulated PBL. Soluble CD23 release was also shown to precede IgE accumulation in the cell-free medium. Addition of compound at later times other than day 0 in the 14 day assay resulted in progressively less inhibition of both IgE and sCD23, and exactly paralleled the effect of an anti-CD23 mAb, MHM6 on IgE levels. Both compounds also inhibited the release of CD23 from human RPMI 8866 cells adoptively transferred i. p. to mice. Doses required for inhibition of CD23 correlated well with the doses required for inhibition of IgE production in IL-4-challenged hu-PBL-SCID mice. IgE was selectively inhibited over total IgG in the SCID mice as well. CONCLUSIONS: Inhibition of CD23 processing alone is sufficient to inhibit IL-4-stimulated IgE production both in vitro and in vivo.

The place of directional coronary atherectomy for the treatment of in-stent restenosis.

The beneficial short and long-term results of coronary stenting have resulted in a dramatic increase in stent utilization, accounting for greater than 80% of coronary interventions [1--9]. However, the long-term beneficial effect of coronary stenting is limited by the occurrence of a 14 to 61% restenosis rate [10--13]. The optimal percutaneous revascularization strategy for the treatment of in-stent restenosis remains undetermined. Although balloon angioplasty has been performed with high initial procedural success, the long-term results are disappointing due to significant recurrence [14--18]. In this article we describe the feasibility, safety, immediate and long-term outcome of directional coronary atherectomy (DCA) as a treatment modality in a cohort of patients undergoing percutaneous intervention for the treatment of in-stent restenosis at the Massachusetts General Hospital.

Mitral balloon valvotomy for patients with mitral stenosis in atrial fibrillation: immediate and long-term results.

OBJECTIVES: The purpose of this study was to examine the effect of atrial fibrillation (AF) on the immediate and long-term outcome of patients undergoing percutaneous mitral balloon valvuloplasty (PMV). BACKGROUND: There is controversy as to whether the presence of AF has a direct negative effect on the outcome after PMV. METHODS: The immediate procedural and the long-term clinical outcome after PMV of 355 patients with AF were prospectively collected and compared with those of 379 patients in normal sinus rhythm (NSR). RESULTS: Patients with AF were older (62 +/- 12 vs. 48 +/- 14 years; p < 0.0001) and presented more frequently with New York Heart Association (NYHA) class IV (18.3% vs. 7.9%; p < 0.0001), echocardiographic score >8 (40.1% vs. 25.1%; p < 0.0001), calcified valves under fluoroscopy (32.4% vs. 18.8%, p < 0.0001) and with history of previous surgical commissurotomy (21.7% vs. 16.4%; p = 0.0002). In patients with AF, PMV resulted in inferior immediate and long-term outcomes, as reflected in a smaller post-PMV mitral valve area (1.7 +/- 0.7 vs. 2 +/- 0.7 cm2; p < 0.0001) and a lower event free survival (freedom of death, redo-PMV and mitral valve surgery) at a mean follow-up time of 60 months (32% vs. 61%; p < 0.0001). In the group of patients in AF, severe post-PMV mitral regurgitation (> or =3+) (p = 0.0001), echocardiographic score >8 (p = 0.004) and pre-PMV NYHA class IV (p = 0.046) were identified as independent predictors of combined events at follow-up. CONCLUSIONS: Patients with AF have a worse immediate and long-term outcomes after PMV. However, the presence of AF by itself does not unfavorably influence the outcome, but is a marker for clinical and morphologic features associated with inferior results after PMV.

Is redo percutaneous mitral balloon valvuloplasty (PMV) indicated in patients with post-PMV mitral restenosis?

OBJECTIVES: The purpose of this study was to assess the immediate and long-term outcome of repeat percutaneous mitral balloon valvuloplasty (PMV) for post-PMV mitral restenosis. BACKGROUND: Symptomatic mitral restenosis develop in 7% to 21% of patients after PMV. Currently, most of these patients are referred for mitral valve replacement. However, it is unknown if these patients may benefit from repeat PMV. METHODS: We report the immediate outcome and long-term clinical follow-up results of 36 patients (mean age 58+/-13 years, 75% women) with symptomatic mitral restenosis after prior PMV, who were treated with a repeat PMV at 34.6+/-28 months after the initial PMV. The mean follow-up period was 30+/-33 months with a maximal follow-up of 10 years. RESULTS: An immediate procedural success was obtained in 75% patients. The overall survival rate was 74%, 72% and 71% at one, two, and three years respectively. The event-free survival rate was 61%, 54% and 47% at one, two, and three years respectively. In the presence of comorbid diseases (cardiac and noncardiac) the two-year event-free survival was reduced to 29% as compared with 86% in patients without comorbid diseases. Cox regression analysis identified the echocardiographic score (p = 0.03), post-PMV mitral valve area (p = 0.003), post-PMV mitral regurgitation grade (p = 0.02) and post-PMV pulmonary artery pressure (p = 0.0001) as independent predictors of event-free survival after repeat PMV. CONCLUSIONS: Repeat PMV for post-PMV mitral restenosis results in good immediate and long-term outcome in patients with low echocardiographic scores and absence of comorbid diseases. Although the results are less favorable in patients with suboptimal characteristics, repeat PMV has a palliative role if the patients are not surgical candidates.

Comparison of immediate and long-term results of mitral balloon valvotomy with the double-balloon versus Inoue techniques.

There is controversy as to whether the double-balloon or Inoue technique of percutaneous mitral balloon valvotomy (PMBV) provides superior immediate and long-term results. This study compares the immediate procedural and long-term outcomes of patients undergoing PMBV using the double-balloon versus the Inoue techniques. Seven hundred thirty-four consecutive patients who underwent PMBV using the double-balloon (n = 621) or Inoue technique (n = 113) were studied. There were no statistically significant differences in baseline clinical and morphologic characteristics between the double-balloon and Inoue patients. The double-balloon technique resulted in superior immediate outcome, as reflected in a larger post-PMBV mitral valve area (1.9 +/- 0.7 vs 1.7 +/- 0.6 cm2; p = 0.005) and a lower incidence of 3+ mitral regurgitation after PMBV (5.4% vs 10.6%; p = 0.05). This superior immediate outcome of the double-balloon technique was observed only in the group of patients with echocardiographic score < or = 8 (post-PMBV mitral valve areas 2.1 +/- 0.7 vs 1.8 +/- 0.6; p = 0.004). Despite the difference in immediate outcome, there were no significant differences in event-free survival at long-term follow-up between the 2 techniques. Our study demonstrates that compared with the Inoue technique, the double-balloon technique results in a larger mitral valve area and less degree of severe mitral regurgitation after PMBV. Despite the difference in immediate outcome between both techniques, there were no significant differences in event-free survival at long-term follow-up.