Human spleen cell generation of factors stimulating human pluripotent stem cell, erythroid, and myeloid progenitor cell growth

Mitogen-stimulated murine spleen cells produce humoral substances capable of supporting murine hematopoiesis and pluripotent stem cell proliferation in vitro. Thus, we evaluated conditioned media generated by human spleen cells (SCM) in the presence or absence of mitogens for factors stimulatory for human pluripotent (CFU-GEMM), erythroid (BFU- E), and myeloid (CFU-GM) precursors. Two and one half percent to 10% SCM stimulated proliferation of all three types of precursor cells from nonadherent buoyant human marrow target cells. Mitogen-stimulated SCM augmented CFU-GM (175% to 225%), whereas CFU-GEMM and BFU-E growth was essentially unchanged. Cell separation procedures used to determine which cells provided these microenvironmental stimuli indicated that nonadherent mononuclear spleen cells provided the bulk of the CSF-GM, whereas adherent cells (95% nonspecific esterase + monocyte- macrophages) and nonadherent cells provided similar proportions of CSF- mix and erythroid burst-promoting activity (BPA). The nonadherent cells generating high levels of CSF-mix, BPA, and CSF-GM were predominantly Leu-1-negative, ie, non-T, cells. In the presence or absence of mitogens, SCM was a more potent source (1.3- to 3.8-fold) than peripheral leukocyte CM of the growth factors for the three progenitor cell types. Specific in situ cytochemical stains for analyzing morphology of myeloid colonies demonstrated that SCM stimulated the proliferation of the same types and proportions of colonies as human placental CM, suggesting that these CMs may contain similar CSF-GMs. These data show the contribution of spleen cell subsets to the generation of hematopoietic growth factors and the responsiveness of these cells to various mitogenic stimuli.     

Pathologic features of reflux and Helicobacter pylori-associated carditis: a comparative study

Inflammation of the gastric cardia, which is the most proximal portion of the stomach, in most instances is the result of either gastroesophageal reflux disease or H. pylori infection. Histologic distinction between these two entities is important because the treatment, natural history, and risk of malignancy are different. Moreover, multilayered epithelium, a possible precursor to Barrett's esophagus, has only recently been described in the gastric cardia, and its relationship to gastroesophageal reflux disease is unknown. The aim of this study was to compare the histologic features of the gastric cardia and the prevalence of multilayered epithelium in patients with reflux versus H. pylori-associated carditis. Routinely processed hematoxylin and eosin-stained mucosal biopsies of the gastric cardia from 30 patients with reflux-associated carditis, 25 with H. pylori-associated carditis, and 30 control patients (no reflux, no H. pylori) were evaluated for a wide variety of histologic features such as goblet cell metaplasia, presence of multilayered epithelium, type of glandular epithelium (mucous, oxyntic, mixed mucous/oxyntic), pancreatic metaplasia, overall degree of inflammation, and the quantity of individual types of inflammatory cells. The clinical and histologic features were compared between the two study groups and controls. Clinically, the reflux carditis group (male/female ratio: 21/9, mean age 56 years) had a significantly higher male/female ratio (p <0.01) and a slightly higher mean age in comparison with the H. pylori group (male/female ratio: 9/16, mean age 50 years). Histologically, the reflux group had significantly less overall inflammation (p <0.05), with fewer plasma cells (p <0.04) and neutrophils (p <0.006), but a higher prevalence of multilayered epithelium [9 of 30 (30%) vs 1 of 25 (4%) in the H. pylori group, p = 0.01]. In the reflux carditis group, multilayered epithelium was significantly associated with neutrophilic inflammation (p <0.05), but not any other features of chronic carditis or with any of the specific epithelial cell types. The control group showed less inflammatory activity in comparison with the H. pylori group and a lower prevalence of multilayered epithelium and eosinophilic inflammation in comparison with the reflux group. The clinical and pathologic features of reflux carditis are distinct from H. pylori carditis and are characterized by less overall inflammation and fewer neutrophils and plasma cells. Multilayered epithelium not uncommonly occurs in the cardia of patients with gastroesophageal reflux disease but without Barrett's esophagus, further supporting our hypothesis that multilayered epithelium may represent an early precursor in the development of columnar metaplasia in Barrett's esophagus.     

Clinical Profile of Special Children at Asha Centre

Background

The care of special children suffering from cerebral palsy, deaf mutism, mental retardation (MR) and post encephalitic sequelae etc. is done in the armed forces at “ASHA” centre supported by Army Wives Welfare Association (AWWA).

Methods

The clinical profile and underlying etiological factors in these children were studied.

Result

Out of 30 children studied, majority were males. The commonest disability was cerebral palsy, seen in 13 (43%) cases followed by mental retardation in six (20%) and post encephalitic sequlae in four (13%) cases. Convulsions were noticed in 12 (40%) cases. The delayed speech was a significant handicap observed in 27 (90%) cases.

Conclusion

Among the etiological factors, natal causes and infections are leading factors in these children and there is an urgent need to strengthen the existing maternal and child health services in our country.Key Words: Cerebral palsy     

Fall-off in Reporting Life Events: Effects of Life Change,Desirability, and Anticipation

Abstract

The influence of event characteristics on recall was examined by directly comparing fall-off in reporting life events as a function of life change, desirability, and anticipation. We collected information from a sample of 1,669 blue-collar workers on stressful life events that occurred in a 1-year interval before the questionnaire was administered. The results indicated no fall-off in reporting events associated with marked life changes (ie, salient events). In contrast, significant fall-off was observed for events characterized by varying degrees of desirability and anticipation. Although ratings of desirability and saliency were not independent, saliency of life events emerged as the dimension most closely associated with accuracy of event reporting. Research on the reliability of measures of life events and the association between event characteristics and illness should consider the kinds of systematic reporting differences observed here.     

Developing Signs of Safety: A Deaf-accessible counselling toolkit for trauma and addiction

The U.S. Deaf community—more than half a million Americans who communicate using American Sign Language (ASL)—experiences higher rates of trauma exposure and substance use disorder (SUD) than the general population. Yet there are no evidence-based treatments for any behavioural health condition that have been evaluated for use with Deaf people. The driving aim of our work, therefore, has been to develop and formally evaluate a Deaf-accessible trauma/SUD counselling approach. Here we describe our initial intervention development work and a single-arm pilot that evaluated the feasibility, acceptability, and preliminary clinical efficacy of Signs of Safety—a Deaf-accessible toolkit to be used with an existing, widely adopted protocol for trauma and addiction (Seeking Safety). Preliminary efficacy results indicated clinically significant reductions in PTSD symptoms and frequency of alcohol use for the Seeking Safety/Signs of Safety model. Frequency of drug use did not change significantly—likely attributable to the mid-study legalization of recreational marijuana in our state. Next steps include the redesign and refilming of Signs of Safety based on pilot participant feedback, again using a Deaf-engaged development and production process. This new toolkit will be tested via a pilot randomized controlled trial designed based on present methodological lessons learned.     

INAUGURAL ARTICLE by a Recently Elected Academy Member:Nonsteroidal anti-inflammatory drug sensitizes Mycobacterium tuberculosis to endogenous and exogenous antimicrobials

Existing drugs are slow to eradicate Mycobacterium tuberculosis (Mtb) in patients and have failed to control tuberculosis globally. One reason may be that host conditions impair Mtb’s replication, reducing its sensitivity to most antiinfectives. We devised a high-throughput screen for compounds that kill Mtb when its replication has been halted by reactive nitrogen intermediates (RNIs), acid, hypoxia, and a fatty acid carbon source. At concentrations routinely achieved in human blood, oxyphenbutazone (OPB), an inexpensive anti-inflammatory drug, was selectively mycobactericidal to nonreplicating (NR) Mtb. Its cidal activity depended on mild acid and was augmented by RNIs and fatty acid. Acid and RNIs fostered OPB’s 4-hydroxylation. The resultant 4-butyl-4-hydroxy-1-(4-hydroxyphenyl)-2-phenylpyrazolidine-3,5-dione (4-OH-OPB) killed both replicating and NR Mtb, including Mtb resistant to standard drugs. 4-OH-OPB depleted flavins and formed covalent adducts with N-acetyl-cysteine and mycothiol. 4-OH-OPB killed Mtb synergistically with oxidants and several antituberculosis drugs. Thus, conditions that block Mtb’s replication modify OPB and enhance its cidal action. Modified OPB kills both replicating and NR Mtb and sensitizes both to host-derived and medicinal antimycobacterial agents.Some bacterial infections can be cured with a single dose of an antibiotic, and most others can be cured with administration of one drug over several days or weeks. In contrast, routine treatment of drug-sensitive tuberculosis (TB) takes 2 mo of therapy with four drugs and an additional 4 mo with two drugs to reduce the 2-y relapse rate below 5%. The difficulty of completing prolonged treatment is a major reason for emergence of drug resistance. When the infecting strain is resistant to isoniazid and rifampin, the two drugs recommended for all 6 mo of treatment, cure often requires 2 y of daily administration of toxic, expensive, second-line agents that are often unavailable at the point of care. When the causative strain is additionally resistant to a quinolone and an aminoglycoside, the resultant “extensively drug-resistant” TB was fatal to 80% of patients in a leading center (1), although complex multidrug regimens have achieved higher cure rates in populations not previously exposed to the additional drugs (2). In addition to sharing air with someone with TB, leading risk factors for contracting the disease are malnutrition, HIV infection, diabetes, and exposure to smoke from cigarettes or cooking fires (3). These epidemiological challenges exacerbate problems of inadequate diagnostic technology and limited access to drug susceptibility testing and to drugs. Control of the pandemic is not in sight (3).It is widely hypothesized that treatment of TB is protracted because nonreplicating (NR) subpopulations of bacilli are phenotypically tolerant to drugs that were selected for activity against replicating (R) Mycobacterium tuberculosis (Mtb) (4). Mtb can occupy diverse microenvironments in the host. Evidence from auxotrophs, analyses of gene expression, and direct and indirect biochemical measurements in vivo or ex vivo in experimental animals and people suggest that such environments expose Mtb to acid, hypoxia, reactive nitrogen intermediates (RNIs), oxidative stress, carbohydrate deficiency, and metal starvation or intoxication, and require Mtb to metabolize fatty acids or cholesterol (5–17). In vitro, many of the same conditions can make Mtb relatively refractory to killing by the standard agents, except for pyrazinamide, which is only effective at a low pH.Thus, there is a need for a high-throughput screen (HTS) for compounds that kill Mtb when the Mtb has been rendered NR by a combination of physiologically relevant host-imposed conditions. We were encouraged to devise such a screen by recent discoveries of a class of compounds that kill Mtb only when it is NR (18), an antibiotic in clinical use for other infections that kills NR Mtb better than R Mtb (19), and a compound that kills NR and R Mtb equally well (20). Unfortunately, only one of those compounds is an approved drug, and even if it were of proven utility in TB, its price would preclude its use by most of those who need it. We decided to screen other existing drugs that are not regarded as antiinfectives for those that kill NR Mtb. Here, we report finding such a drug in an HTS that combined four host-imposed conditions, some of which converted the drug into a form active on both R and NR Mtb.     

Antibiotic effects on bacterial profile in osteonecrosis of the jaw

Oral Diseases (2011) 18 , 85–95 Objective: Oral infection is considered to play a critical role in the pathogenesis of bisphosphonate‐related osteonecrosis of the jaw (BRONJ), and antibiotic therapy has become a mainstay of BRONJ therapy. This study was aimed to investigate the effect of antibiotics on bacterial diversity in BRONJ tissues. Materials and methods: The bacterial profile from soft tissues associated with the BRONJ lesion was determined using 16S rRNA‐based denaturing gradient gel electrophoresis (DGGE) and sequencing. Twenty BRONJ subjects classified as stage 0–2 were enrolled in this study, and patient groups were divided into an antibiotic cohort (n = 10) treated with systemic antibiotic and a non‐antibiotic cohort (n = 10) with no prior antibiotic therapy. Results: The DGGE fingerprints indicated no significant differences in bacterial diversity of BRONJ tissue samples. Patients on antibiotics had higher relative abundance of phylum Firmicutes with bacterial species, Streptococcus intermedius, Lactobacillus gasseri, Mogibacterium timidum, and Solobacterium moorei, whereas patients without antibiotics had greater amounts of Parvimonas micra and Streptococcus anginosus. Thirty percent of bacterial populations were uncultured (yet‐to be cultured) phylotypes. Conclusion: This study using limited sample size indicated that oral antibiotic therapy may have a limited efficacy on the bacterial population associated with BRONJ lesions.