Measuring abscission spatiotemporal dynamics using quantitative high-resolution microscopy.

The spatiotemporal characteristics of ESCRT (Endosomal Sorting Complex Required for Transport)-mediated mammalian cytokinetic abscission have been studied in recent years using quantitative high-resolution light microscopy techniques. Here we describe how to apply spinning disk live cell imaging and structured illumination microscopy (SIM) to define the dynamics and structural organization of abscission and of proteins involved in abscission in a quantitative manner. We further provide a protocol to correlate the structural data, obtained by SIM, to the dynamic information obtained by live cell recordings.

Some puzzling situations in the onset, occurrence and future of coronary heart disease in developed and developing populations, particularly such in sub-Saharan Africa.

Coronary heart disease (CHD) was rare in developed populations until the early 1900s; this prevailed even among the small segments who were prosperous and who, in measure, had most of the currently recognised risk factors. However, in the 1930s, with improved circumstances from general rises in socio-economic state, there were major increases in the occurrence and mortality rate from the disease, the latter reaching a third of the total mortality in some countries, as in the United Kingdom (UK). Puzzlingly, the inter-population diversity of the increases in CHD has been such that there are as much as five fold differences in CHD mortality rates, as, for example, between Poland and Spain. Within recent years, with appropriate treatments, the mortality rate has halved in some countries, again, as in the UK. However, the incidence rate of the disease has diminished little or hardly at all. Risk factors include a familial component and, nutritionally, over-eating, a high fat intake, relatively low intakes of plant foods, especially of vegetables and fruit and, non-nutritionally, smoking, excessive alcohol consumption and a low level of everyday physical activity. On the one hand, known risk factors, broadly, are considered to be capable of explaining only about half of the variation in the occurrence of the disease. Even at present, known risk factors far from fully explain the epidemiological differences in mortality rates. Yet, on the other hand, there is abundant evidence that in population groups, among whom risk factors are low or have been reduced, CHD incidence and mortality rates are lower. Notwithstanding this knowledge, broadly, there is very little interest in the general public in taking avoiding measures. As to the situation in developing populations, in sub-Saharan Africa, in urban Africans, as in Johannesburg, South Africa, despite considerable westernisation of life style and with rises in risk factors, CHD remains of very low occurrence, the situation thereby resembling, historically, its relatively slow emergence in developed populations. In most eastern countries, mortality rates remain relatively low, as in Russia and Japan. However, in major contrast, in India, rates have risen considerably in urban dwellers. Indeed, in Indian immigrants, as in those in the UK, their rate actually exceeds that in the country's white population. In brief, much remains to be explained in the epidemiology of the disease.

Major differences in the IgG subclass response to Helicobacter pylori in the first and third worlds.

BACKGROUND: Studies in developed countries would suggest that the immune response to Helicobacter pylori infection is a T helper cell I predominant response. Unlike subjects from developed countries, those resident in developing countries are subject to infection with a myriad of gastrointestinal pathogens from early in life. Given that H. pylori is acquired early in life, such infections may alter the immune response to H. pylori. The aim of this study was to compare the immune response to H. pylori in subjects from developed and developing countries. METHODS: Using a previously validated IgG subclass ELISA, the H. pylori specific IgG I/IgG2 subclass ratio (a marker of the T helper cell response) in 58 adult and 21 paediatric symptomatic H. pylori positive Sowetan subjects was compared with that in 64 Australian and 45 German symptomatic H. pylori positive subjects. RESULTS: An IgGI predominant response (IgG1/IgG2 ratio >1) was observed in 81% of Sowetan adults and 90% of children compared with 4.7% of Australians and 4.4% of Germans. The IgG1/IgG2 ratio was significantly higher in Sowetans compared with Australians and Germans (P < 0.001). In Australian and German subjects the IgG1/IgG2 ratio was significantly higher in NUD compared with DU. No significant difference was observed between NUD and other disease states in Sowetans. CONCLUSIONS: This study is the first to provide evidence that the host immune response to H. pylori infection in an African population differs to that observed in subjects from developed countries. Further studies are required to determine if this occurs in other developing countries.

Colorectal cancer in an African city population in transition.

An enquiry has been made on a series of African patients with colorectal cancer who were admitted in 1995-1999 to Chris Hani Baragwanath Hospital (3200 beds), and who lived in Soweto (population about 1 million), Johannesburg, South Africa. In the urban context described, Africans have considerably more advantages, socio-economically, dietarily and in other respects, than their rural counterparts. The 126 patients comprised 58 males and 68 females, giving calculated incidence rates in their communities for colorectal cancer of 1.7 and 2.0, respectively, per 100,000 'world' population. In contrast, as indicated in the South African Cancer Registry for 1993-1995, the corresponding rates for white males and females were 24.7 and 19.3, respectively, per 100,000. The proportion of African patients under 40 years was 19.0%; but was only 4.0% in the white population. In contrast to this major disparity, there was only a minor interethnic disparity regarding cancers that are very common in Africans, namely, those of the oesophagus and lung. Hence, with ongoing transitional changes - in diet and other respects - the relatively high proportion of younger African patients probably indicates a rising occurrence of colorectal cancer in the urban African population.

Tropical enteropathy: a T-cell-mediated crypt hyperplastic enteropathy.

OBJECTIVE: Tropical enteropathy is widespread throughout the tropics, but its pathogenesis is unknown. T-cell activation has been demonstrated to result in enteropathy in vitro and in animal models, and occurs in untreated patients with coeliac disease. We have therefore examined the hypothesis that T-cell activation is important in the pathogenesis of tropical enteropathy. PATIENTS AND METHODS: Healthy black Zambian subjects were compared with black and white South Africans. Quantitative microscopy was conducted on distal duodenal biopsies. Mucosal T-cell activation was quantitated by dual colour immunofluorescence staining for CD3 plus CD69 or HLA-DR. Crypt proliferation was measured by direct counting of Feulgen-stained mitotic figures, and systemic immune activation by assay of serum tumour necrosis factor alpha (TNF-alpha). RESULTS: Villous height was reduced (P = 0.0004), crypt depth increased (P < 0.0001), and mitoses per crypt increased (P = 0.014) in black Zambians compared with black and white South Africans. Mucosal thickness was similar. Intraepithelial lymphocyte count was increased in the black groups compared with whites (P = 0.03). CD3+CD69+ (P = 0.0007) and CD3+HLA-DR+ (P < 0.0001) expression was increased in black Zambians compared with black and white South Africans. Serum TNF-alpha was similar in all groups. CONCLUSIONS: Tropical enteropathy is associated with mucosal T-cell activation and crypt hyperplasia. Tropical enteropathy occurs in the absence of malnutrition, diarrhoea or systemic illness.

Helicobacter pylori--an African perspective.

Helicobacter pylori is ubiquitous in Africa, with acquisition in childhood the rule. Despite the prevalence of a virulent strain (in Soweto, most H. pylori organisms are cagA- and vacAS(1)-positive) H. pylori-associated pathology (duodenal ulcer, gastric ulcer and gastric cancer) has a variable, often low distribution in sub-Saharan Africa that does not parallel H. pylori prevalence in the population, suggesting a different natural history from that seen in developed countries. Progression to atrophic gastritis in Africans does not appear to differ from that reported in other regions, but as yet unidentified factors may play a role in inhibiting progression to gastric cancer. Studies have suggested that the specific IgG subclass response to H. pylori is predominately IgG1 (suggestive of a Th2 response), and the Th2 response may provide a protective effect against development of gastric cancer. Host immune mechanisms may be the key to different responses to H. pylori in the developed and developing worlds.

The gastro-oesophageal reflux disease complex in sub-Saharan Africa.

Epidemiological and clinical studies that have reported on gastro-oesophageal reflux disease (GERD), Barrett's oesophagus, oesophageal adenocarcinoma and Helicobacter pylori infection in sub-Saharan Africa were reviewed. The data indicate that Barrett's oesophagus is rare and oesophageal adenocarcinoma uncommon in all regions of sub-Saharan Africa studied (South Africa, Ethiopia, Nigeria, Zimbabwe, Kenya and Uganda). Hiatus hernia is also uncommon. There are too few reports of GERD to allow comment. The overwhelming majority of oesophageal cancers are squamous cell type. H. pylori infection is ubiquitous with an overall prevalence of 61-100%. It is concluded that although urbanization has resulted in an increase of risk factors associated with GERD, which would be expected to lead to an increase in this disease among Africans, this increase has not happened. It is believed that the critical factor preventing GERD in black Africans is H. pylori infection, which is usually acquired in childhood, is lifelong and is probably protective for the oesophagus.

Gastric cancer in sub-Saharan Africa.

Gastric cancer has a variable but generally low prevalence in black populations of sub-Saharan Africa, despite a high prevalence of Helicobacter pylori (the 'African enigma'). Evidence from Soweto indicates that the host response to H. pylori may be protective against a virulent organism and that, in most people, H. pylori does not lead to more serious sequelae. This suggests that there may be host protective/inhibitory factors present, which prevent the progression of H. pylori-induced chronic active gastritis to cancer.

Hepatitis B and C virus infections and liver function in AIDS patients at Chris Hani Baragwanath Hospital, Johannesburg.

BACKGROUND: Impaired liver function tests and co-infection with hepatitis viruses in AIDS patients are common in western countries. OBJECTIVE: To assess liver function and prevalence of co-infection with hepatitis B and hepatitis C viruses in AIDS patients at Chris Hani Baragwanath Hospital. DESIGN: A prospective study. SETTING: Chris Hani Baragwanath Hospital, Johannesburg, South Africa. PATIENTS: One hundred consecutive patients with AIDS admitted to Chris Hani Baragwanath Hospital. RESULTS: There were 52 males and 48 females aged 16 to 54 years (mean + SD: 34.6 + 7.5 years). The results of laboratory test were as follows: LFTs: bilirubin 11.8 (+15.6) mumol/l; AST: 79.6 (+/- 116.6) iu/L; alkaline phosphatase: 204.3 (+/- 237.4) i mu/L; albumin: 23.9 (+/- 6.2) g/l; CD4+ lymphocytes: 141.5 (+/- 168.6) microliters; CD8+: 666.9 (+/- 618.3) microliters; HBV - HbsAg: 6 (6%); HbsAg + eAg: 3 (3%); previous disease (Anti HBs and/or anti HBc): 35%, HCV: 1(1%). CONCLUSION: Liver function tests were impaired in the majority of patients with AIDS (93%) in our setting. Evidence of previous and present HBV infection was present in 41%. This is different from what is observed in western countries (90-95%). The results also suggest that patients here acquired HBV infection while still immuno competent. HCV infection was rare.

Sequential inactivation of rdxA (HP0954) and frxA (HP0642) nitroreductase genes causes moderate and high-level metronidazole resistance in Helicobacter pylori.

Helicobacter pylori is a human-pathogenic bacterial species that is subdivided geographically, with different genotypes predominating in different parts of the world. Here we test and extend an earlier conclusion that metronidazole (Mtz) resistance is due to mutation in rdxA (HP0954), which encodes a nitroreductase that converts Mtz from prodrug to bactericidal agent. We found that (i) rdxA genes PCR amplified from 50 representative Mtz(r) strains from previously unstudied populations in Asia, South Africa, Europe, and the Americas could, in each case, transform Mtz(s) H. pylori to Mtz(r); (ii) Mtz(r) mutant derivatives of a cultured Mtz(s) strain resulted from mutation in rdxA; and (iii) transformation of Mtz(s) strains with rdxA-null alleles usually resulted in moderate level Mtz resistance (16 microg/ml). However, resistance to higher Mtz levels was common among clinical isolates, a result that implicates at least one additional gene. Expression in Escherichia coli of frxA (HP0642; flavin oxidoreductase), an rdxA paralog, made this normally resistant species Mtz(s), and frxA inactivation enhanced Mtz resistance in rdxA-deficient cells but had little effect on the Mtz susceptibility of rdxA(+) cells. Strains carrying frxA-null and rdxA-null alleles could mutate to even higher resistance, a result implicating one or more additional genes in residual Mtz susceptibility and hyperresistance. We conclude that most Mtz resistance in H. pylori depends on rdxA inactivation, that mutations in frxA can enhance resistance, and that genes that confer Mtz resistance without rdxA inactivation are rare or nonexistent in H. pylori populations.

Differences in genotypes of Helicobacter pylori from different human populations.

DNA motifs at several informative loci in more than 500 strains of Helicobacter pylori from five continents were studied by PCR and sequencing to gain insights into the evolution of this gastric pathogen. Five types of deletion, insertion, and substitution motifs were found at the right end of the H. pylori cag pathogenicity island. Of the three most common motifs, type I predominated in Spaniards, native Peruvians, and Guatemalan Ladinos (mixed Amerindian-European ancestry) and also in native Africans and U.S. residents; type II predominated among Japanese and Chinese; and type III predominated in Indians from Calcutta. Sequences in the cagA gene and in vacAm1 type alleles of the vacuolating cytotoxin gene (vacA) of strains from native Peruvians were also more like those from Spaniards than those from Asians. These indications of relatedness of Latin American and Spanish strains, despite the closer genetic relatedness of Amerindian and Asian people themselves, lead us to suggest that H. pylori may have been brought to the New World by European conquerors and colonists about 500 years ago. This thinking, in turn, suggests that H. pylori infection might have become widespread in people quite recently in human evolution.

Fermentation of dietary starch in humans.

OBJECTIVE: Dietary starch that escapes digestion in the small intestine may be quantitatively more important than dietary fiber as a substrate for fermentation. The products of fermentation have important implications in the pathogenesis of colorectal cancer and other diseases of the large bowel, which are uncommon in Africans but have a high prevalence in Western populations. METHODS: Maize porridge is a staple of most blacks in South Africa. Stale maize porridge (high-resistant starch [HRS]) seems to induce greater fermentation in the large bowel than fresh maize porridge (low-resistant starch [LRS]). RESULTS: In the present study, healthy colostomy subjects fed stale maize porridge had significantly more production of short-chain fatty acids (SCFA) (mean SCFA, HRS = 182.6; mean SCFA, LRS = 116.1; p < 0.05) in their colostomy effluent together with a significant drop in stool pH (mean pH, HRS = 5.91; mean pH, LRS = 6.70; p < 0.001). The SCFA butyrate (mean, HRS = 35.1; mean, LRS = 17.6; p < 0.05) and acetate (mean, HRS = 93.9; mean, LRS = 65.8; p < 0.05) were significantly elevated on the stale maize porridge diet when compared with consumption of fresh maize porridge. SCFA propionate (mean, HRS = 43.1; mean, LRS = 24.8; p = 0.05), also increased with stale maize porridge, but was not statistically significant. CONCLUSION: A high-resistant starch diet and its resultant increase in fermentation products may be partly responsible for protecting the black population against colorectal cancers and other large bowel diseases.

Dendritic spine formation and pruning: common cellular mechanisms?

The recent advent of novel high-resolution imaging methods has created a flurry of exciting observations that address a century-old question: what are biological signals that regulate formation and elimination of dendritic spines? Contrary to the traditional belief that the spine is a stable storage site of long-term neuronal memory, the emerging picture is of a dynamic structure that can undergo fast morphological variations. Recent conflicting reports on the regulation of spine morphology lead to the proposal of a unifying hypothesis for a common mechanism involving changes in postsynaptic intracellular Ca2+ concentration, [Ca2+]i: a moderate rise in [Ca2+]i causes elongation of dendritic spines, while a very large increase in [Ca2+]i causes fast shrinkage and eventual collapse of spines. This hypothesis provides a parsimonious explanation for conflicting reports on activity-dependent changes in dendritic spine morphology, and might link these changes to functional plasticity in central neurons.

Temporal analysis of connexin43 protein and gene expression throughout the menstrual cycle in human endometrium.

OBJECTIVE: To analyze the pattern of connexin43 gene and protein expression in human endometrium throughout the menstrual cycle. DESIGN: Controlled clinical study. SETTING: An academic research center. PATIENT(S): Women with 28-day menstrual cycles who had mechanical infertility and failed to conceive after IVF treatment. INTERVENTION(S): Endometrial and blood samples were collected on days 8, 12, 14, 21, and 25 of spontaneous menstrual cycles. MAIN OUTCOME MEASURE(S): Endometrial expression of connexin43 protein and messenger RNA, endometrial thickness, and serum concentrations of gonadotropins and steroids. RESULT(S): The expression of connexin43 gene and protein decreased on day 12 and day 14 of the menstrual cycle and then increased on day 21 and day 25, respectively. A serum LH surge accompanied by a peak in the FSH concentration was observed on days 12-14. The progesterone concentration increased on days 21-25, but there was no significant change in the E2 concentration. The thickness of the endometrium increased between days 8 and 12 and did not change further between days 21 and 25. CONCLUSION(S): The expression of connexin43 gene and protein in human endometrium changes during the menstrual cycle in a pattern that is associated with the secretion of LH, FSH, and progesterone. This pattern may serve as a marker for implantation competence.

Case control study of environmental factors in the etiology of the first attack of acute pancreatitis: a pilot study.

The aim of this case control study was to assess environmental factors in the etiology of the first attack of acute pancreatitis (AP) in Soweto, South Africa, in the light of modern developments. The study group consisted of 30 patients presenting with a first attack of AP, and 30 healthy age- and gender-matched controls. Fruit intake was found to be a protective factor (odds ratio [OR] 5.3). Crude ORs, although of marginal importance, showed that daily alcohol intake and years of exposure to occupational chemicals may be of significance but would require a much larger study to test these factors. This would be necessary in order to explain the rapid increase in pancreatitis coincident with urbanization and industrialization in the community.

Is redshift-dependent evolution of galaxies a theoretical artifact?

The physical validity of the hypothesis of (redshift-dependent) luminosity evolution in galaxies is tested by statistical analysis of an intensively studied complete high-redshift sample of normal galaxies. The necessity of the evolution hypothesis in the frame of big-bang cosmology is confirmed at a high level of statistical significance; however, this evolution is quantitatively just as predicted by chronometric cosmology, in which there is no such evolution. Since there is no direct observational means to establish the evolution postulated in big-bang studies of higher-redshift galaxies, and the chronometric predictions involve no adjustable parameters (in contrast to the two in big-bang cosmology), the hypothesized evolution appears from the standpoint of conservative scientific methodology as a possible theoretical artifact.