Associations of stunting at 2 years with body composition and blood pressure at 8 years of age: longitudinal cohort analysis from lowland Nepal.

BACKGROUND: Stunting remains a very common form of child malnutrition worldwide, particularly in South Asian populations. There is poor understanding of how it develops and how it is associated with subsequent phenotype. SUBJECTS/METHODS: We used data from a longitudinal cohort of children (n = 841) in lowland Nepal to investigate associations of stunting at 2 years with maternal traits and early growth patterns, and with body size and composition, kidney dimensions by ultrasound, lung function by spirometry and blood pressure (BP) at 8 years. RESULTS: Compared to non-stunted children, children stunted at 2 years came from poorer families and had shorter, lighter mothers. They tended to have higher birth order, were born smaller, and remained shorter, lighter and thinner at 8 years. They had lower leg length, lean and fat masses, smaller kidneys, and reduced lung function (all p < 0.0001). These differences persisted with smaller magnitude after adjusting for current height, maternal height and education, family assets and birth order. Stunting was not associated with BP. DISCUSSION: Stunting developed on an inter-generational timescale in this population and its risk increased with birth order. At 8 years, children stunted at 2 years had deficits in tissue masses and some aspects of physical function that were only partially attributable to their persisting short height and maternal phenotype. This suggests that the early stunting is associated with greater deficits in long-term outcomes than would be expected from the persistent short stature alone.

Effectiveness of female community health volunteers in the detection and management of low-birth-weight in Nepal.

INTRODCTION: Low birth weight (LBW) is a major risk factor for neonatal death. However, most neonates in low-income countries are not weighed at birth. This results in many LBW infants being overlooked. Female community health volunteers (FCHVs) in Nepal are non-health professionals who are living in local communities and have already worked in a field of reproductive and child health under the government of Nepal for more than 20 years. The effectiveness of involving FCHVs to detect LBW infants and to initiate prompt action for their care was studied in rural areas of Nepal. METHODS: FCHVs were tasked with weighing all neonates born in selected areas using color-coded spring scales. Supervisors repeated each weighing using electronic scales as the gold standard comparator. Data on the relative birth sizes of the infants, as assessed by their mothers, were also collected and compared with the measured weights. Each of the 205 FCHVs involved in the study was asked about the steps that she would take when she came across a LBW infant, and knowledge of zeroing a spring scale was also assessed through individual interviews. The effect of the background social characteristics of the FCHVs on their performance was examined by logistic regression. This study was nested within a community-based neonatal sepsis-management intervention surveillance system, which facilitated an assessment of the performance of the FCHVs in weighing neonates, coverage of FCHVs' visits, and weighing of babies through maternal interviews. RESULTS: A total of 462 babies were weighed, using both spring scales and electronic scales, within 72 hours of birth. The prevalence of LBW, as assessed by the gold standard method, was 28%. The sensitivity of detection of LBW by FCHVs was 89%, whereas the sensitivity of the mothers' perception of size at birth was only 40%. Of the 205 FCHVs participating in the study, 70% of FCHVs understood what they should do when they identified LBW and very low birth weight (VLBW) infants. Ninety-six per cent could describe how to zero a scale and approximately 50% could do it correctly. Seventy-seven per cent of FCHVs weighed infants at least once during the study period, and 19 of them (12%) miscategorized infant weights. Differences were not detected between the background social characteristics of FCHVs who miscategorized infants and those who did not. On the basis of maternal reporting, 67% of FCHVs who visited infants had weighed them. CONCLUSIONS: FCHVs are able to correctly identify LBW and VLBW infants using spring scales and describe the correct steps to take after identification of these infants. Use of FCHVs as newborn care providers allows for utilization of their logistical, geographical, and cultural strengths, particularly a high level of access to neonates, that can complement the Nepalese healthcare system. Providing additional training to and increasing supervision of local FCHVs regarding birth weight measurement will increase the identification of high-risk neonates in resource-limited settings.

Long-term melatonin administration attenuates low-LET gamma-radiation-induced lymphatic tissue injury during the reproductively active and inactive phases of Indian palm squirrels (Funambulus pennanti).

A comparative analysis of low linear energy transfer (LET) gamma-radiation-induced damage in the lymphatic tissue of a tropical seasonal breeder, Indian palm squirrel (Funambulus pennanti), during its reproductively active phase (RAP) and inactive phase (RIP) was performed with simultaneous investigation of the effects of long-term melatonin pre-treatment (100 microg/100 g body weight). A total of 120 squirrels (60 during RAP and 60 during RIP) were divided into 12 groups and sacrificed at 4, 24, 48, 72 and 168 h following 5 Gy gamma-radiation exposure; control groups were excluded from exposure. Total leukocyte count and absolute lymphocyte count (ALC) and melatonin only of peripheral blood, stimulation index, thiobarbituric-acid-reactive substances (TBARS) level, superoxide dismutase (SOD) activity, and the apoptotic index of spleen as analysed by terminal deoxynucleotidyl transferase (TdT)-mediated deoxyuridine triphosphate (dUTP) nick-end labelling (TUNEL) noted at observed time-points were significantly reduced in melatonin pre-treated groups during RAP and RIP. Long-term melatonin pre-treatment mitigated radiation-induced alterations more prominently during RIP, as assessed by ALC, TBARS, SOD, TUNEL and caspase-3 activity, at some time-points. Our results demonstrate an inhibitory role of melatonin on caspase-3 activity in splenocytes during RAP and RIP following gamma-radiation-induced caspase-mediated apoptosis. Hence, we propose that melatonin might preserve the viability of immune cells of a seasonal breeder against background radiation, which is constantly present in the environment.

Extracellular calcium protects against verapamil-induced metaphase-II arrest and initiation of apoptosis in aged rat eggs.

Non-specific L-type calcium channel blockers, such as verapamil (> or =50 microM), induce metaphase-II (M-II) arrest and apoptosis in aged rat eggs cultured in Ca(2+)-deficient medium. However, the effects of extracellular Ca(2+) on verapamil-induced M-II arrest and apoptosis have not yet been reported. We have demonstrated that postovulatory aging induced exit from M-II arrest by extruding a second polar body, a morphological sign of spontaneous egg activation (SEA). Verapamil inhibited SEA and induced egg apoptosis in a dose-dependent manner in Ca(2+)-deficient medium. The initiation of apoptotic features was observed at 50 microM of verapamil. Extracellular Ca(2+) (1.80 mM) reduced intracellular H2O2 level, bax protein expression, caspase-3 activity, DNA fragmentation and protected against 50 microM, but not higher concentrations of > or =100 microM in verapamil-induced egg apoptosis. These results suggest that extracellular Ca(2+) ions have a role during SEA and protect against verapamil induced apoptosis in aged rat eggs.

Intracellular levels of hydrogen peroxide and nitric oxide in oocytes at various stages of meiotic cell cycle and apoptosis.

The objective was to find out the functional roles of hydrogen peroxide (H(2)O(2)) and nitric oxide (NO) during various stages of meiotic cell cycle and apoptosis in rat oocytes. For this purpose, 30 oocytes from each stage such as diplotene, metaphase-I (M-I), metaphase-II (M-II) and apoptosis were collected and intracellular H(2)O(2), total nitrite level and inducible nitric oxide synthase (iNOS) expression were analysed. This study demonstrated that generation of a tonic level of H(2)O(2) induces meiotic resumption in diplotene-arrested oocytes and further increase may lead to apoptosis. Conversely, reduction in iNOS expression and total nitrite level are associated with meiotic resumption in diplotene-arrested oocytes, but induce apoptosis in aged oocytes. These results suggest that generation of a tonic level of H(2)O(2), reduced iNOS expression and total nitrite level are associated with meiotic resumption, while more generation of H(2)O(2) and sustained reduced total nitrite level are linked with oocyte apoptosis in rat.

Internal hernia through an iatrogenic defect in the falciform ligament: a case report.

The incidence of internal hernia through a defect in the falciform ligament, mostly congenital, is very rare. In this era of minimally invasive laparoscopic surgeries, a few cases of internal hernia through an iatrogenic defect in the falciform ligament have also been reported. Here, we present a case of a 65-year-old patient who presented with acute small-bowel obstruction. The patient had undergone a laparoscopic fundoplication 4 years ago. On diagnostic laparoscopy, it was found that the cause of the intestinal obstruction was herniation of the small bowel through a window in the falciform ligament (which was probably created due to port insertion during the previous surgery of laparoscopic fundoplication). The obstruction was relieved by the division of the falciform ligament.

Calcium ionophore-induced egg activation and apoptosis are associated with the generation of intracellular hydrogen peroxide.

The present study was designed to investigate whether calcium ionophore-induced activation and apoptosis are associated with the generation of hydrogen peroxide (H(2)O(2)) in rat eggs cultured in vitro. Culture of metaphase-II (M-II) arrested eggs in Ca(2+)/Mg(2+)-deficient medium did not induce egg activation, while a second polar body was observed in 20% of eggs when cultured in Ca(2+)/Mg(2+)-supplemented medium. In Ca(2+)/Mg(2+)-deficient medium, lower concentrations of calcium ionophore (0.2,0.4 and 0.8 microm) not only induced egg activation in a dose-dependent manner but also generation of intracellular H(2)O(2) (84.40+/-0.50 ng/egg) when compared to control eggs (80.46+/-1.34 ng/egg). The higher concentration of calcium ionophore (1.6 microm) induced apoptosis and pronounced generation of intracellular H(2)O(2) (92.43+/-0.93 ng/egg) in treated eggs. Conversely, cell-permeant antioxidant such as 2(3)-tert-butyl-4-hydroxyanisole (BHA) reduced intracellular H(2)O(2) level (81.20+/-1.42 ng/egg) and protected against calcium ionophore-induced morphological changes characteristics of egg activation and apoptosis. These results clearly suggest that calcium ionophore-induced activation and apoptosis are associated with the generation of intracellular H(2)O(2) in rat eggs.

Hydrogen peroxide modulates meiotic cell cycle and induces morphological features characteristic of apoptosis in rat oocytes cultured in vitro.

Hydrogen peroxide (H2O2) is known to induce cell cycle arrest and apoptosis in various somatic cell types cultured in vitro. We hypothesize that this reactive oxygen species (ROS) could modulate cell cycle and induce morphological features characteristics of apoptosis in oocytes cultured in vitro. To test this hypothesis, immature and mature oocytes were cultured in medium containing various doses of H2O2 with or without caspase-3 inhibitor for various times. The treatment of H2O2 induced germinal vesicle break down (GVBD) in all immature oocytes followed by initiation of shrinkage. Some of immature oocytes (but not mature oocytes) also showed membrane blebbing. On the other hand, H2O2 treatment inhibited first polar body emission in mature oocytes just prior to initiation of shrinkage. The cytoplasmic granulation and fragmentation into apoptotic bodies were observed in mature oocytes during later stages of H2O2 treatment. The shrinkage was induced by H2O2 in a dose- and time-dependent manner in both immature and mature oocytes. Although, H2O2-induced degeneration was observed in both immature and mature oocytes after 2.0 hrs of treatment, immature oocytes were more susceptible to undergo quick shrinkage, membrane blebbing and degeneration. Co-addition of caspase-3 inhibitor prevented shrinkage and degeneration of both immature and mature oocytes except membrane blebbing that was observed at higher doses of H2O2 after 1.0 hr of culture. Treatment of H2O2 induced bax protein expression (3 times), DNA fragmentation and caspase-3 activity (2.5 times) in oocytes undergoing morphological apoptotic changes. These findings clearly suggest that H2O2 induced GVBD in immature oocytes, inhibited first polar body extrusion in mature oocytes prior to initiation of morphological changes characteristic of apoptosis such as shrinkage, membrane blebbing and cytoplasmic fragmentation prior to degeneration.

Effect of styrene maleic anhydride (SMA) on viability and integrity of isolated rat oocytes in vitro.

In vitro spermicidal property of the polymer, styrene maleic anhydride (SMA), a compound under clinical trial as an intravasal male contraceptive, is already established. The present study was designed to evaluate the effect of the polymer (SMA) on the viability and integrity of rat oocytes in vitro. The pH-lowering effect of this polymer was evident following precipitation in Medium 199. Oocytes became nonviable with clearly resolved granulated cytoplasm following incubation with undiluted polymeric first wash medium. Dilution of polymeric wash medium prolonged the integrity and viability of oocytes under incubation. The potency of the first wash medium in terms of affecting the viability of oocytes was highest among different washes and dilutions. Bulging of cytoplasm and an enlarged zona pellucida envelope were observed in a few oocytes treated with the first wash. These findings have shown that, like sperm, oocytes also lose their integrity and viability when coincubated with the polymer or medium containing the polymer. However, developing an appropriate technology for delivery is crucial to exploit this potency of the polymer for development of a female contraceptive.

Effect of cyclosporine on human sperm motility in vitro.

Cyclosporine affects motility and viability of human sperm when incubated together in vitro. Sperm motility was almost reduced to nil following 10 min of incubation with cyclosporine at a concentration of 1 mg/mL. However, 200 micrograms/mL of the drug has no effect on motility and viability when tested for up to 60 min under standard laboratory conditions. Cyclosporine effect on sperm was both dose and time dependent. Sperm sensitivity and susceptibility to cyclosporine even to lower doses increased significantly following withdrawal of bovine serum albumin from the incubating medium. Compared to untreated controls, lactate dehydrogenase was estimated higher by more than 2 to 4 times in the sperm-free incubating media, suggesting an altered membrane porosity in the affected spermatozoa.

The in vitro effects of forskolin, IBMX and cyanoketone on meiotic maturation in follicle-enclosed catfish (Clarias batrachus) oocytes.

The effects of an adenylate cyclase activator (forskolin, FK), phosphodiesterase inhibitor (3-isobutyl-l-methyl-xanthine, IBMX) and an inhibitor of steroidogenesis (cyanoketone, CK) on germinal vesicle breakdown (GVBD) in the catfish (Clarias batrachus) were investigated in vitro. In most of the experiments GVBD was induced by using 1 microgram/ml 17 alpha,20 beta-dihydroxy-4-pregnen-3-one (17 alpha,20 beta-DP), which is the maturation-inducing steroid (MIS) for this species. Adenosine 3':5'-cyclic monophosphate (cAMP) levels were also measured in the control, MIS-induced and/or FK- and IBMX-treated follicle-enclosed oocytes. MIS-induced GVBD was inhibited by FK (> or = 0.5 microM) or IBMX (> or = 1.0 mM), but oocyte exposed to 0.1 microM FK or 0.5 mM IBMX, after MIS stimulation, underwent GVBD. However, an inhibition of GVBD was recorded when the MIS-induced folliculated oocytes were preincubated with CK (1 microgram/ml) and subsequently treated with 0.1 microM FK. In the time course study, when the oocytes were stimulated by MIS for various time intervals and then treated with 1.0 microM FK or 1.0 mM IBMX, both the substances blocked maturation if they were added up to 12 hr after MIS. The extent of inhibition was gradually decreased and was completely removed after 30 hr of post-MIS stimulation. The stimulatory dose of 17 alpha,20 beta-DP (1 microgram/ml) not only induced GVBD (83.2 +/- 1.50%) in vitro but also reduced oocyte cAMP level (65.3 +/- 2.85 pmol/100 micrograms protein) significantly after 6 hr of incubation. However, FK (10.0 microM) or IBMX (1.0 mM) countered these effects and promoted the accumulation of cAMP in the oocytes; FK being more potent. On the other hand, when unstimulated full-grown but immature oocytes were cultured in vitro in the presence of different concentrations of FK, an induction of oocyte maturation was recorded in dose- and time-dependent manner. These results strongly suggest the involvement of cAMP in the regulation of catfish oocyte maturation.

Urogenital anomalies in fetal rats produced by the anticancer agent 4(5)-(3,3-dimethyl-1-triazeno) imidazole-4-carboxamide.

Pregnant Wistar rats injected intraperitoneally on gestational day 12 with single doses (100-1,000 mg/kg) or 600 mg/kg of 4(5)-(3,3-dimethyl-1-triazeno) imidazole-4-carboxamide (dic) were autopsied on day 21 (100-1,000 mg/kg) or at 24-hour intervals on days 13-20 (600 mg/kg). Controls received CMC on the same schedule. All fetuses were weighed and examined for urogenital system (UGS) malformations. Those given 600 mg/kg were also studied histologically. DIC produced significant growth retardation at all doses on day 21 (18-72%). UGS malformations occurred in 27-67% of the fetuses at 200-400 mg/kg and in 100% of those given 600 mg/kg or more of DIC. Abnormalities included renal growth inhibition, fusion, ectopia, and ureteropelvic dilatation. At 600 mg/kg renal and body weights were reduced 40 and 55%, respectively. Ureteropelvic dilation was common, and cortical glomeruli, nephric collecting tubules, and papillae were retarded in development. The juxtamedullary glomeruli were well developed. Proximal nephric tubular mitotic activity was 85% greater than in control animals (day 17). On the basis of pertinent morphological and physiological data, it is postulated that the dilated upper urinary tracts represent functional hydronephrosis incident to severe renal retardation and its resultant compensatory response.

Cellular and biochemical aspects of growth retardation in rat fetuses induced by maternal administration of selected anticancer agents.

Single ip injections of 600 mg/kg 5-(3,3-dimethyl-1-triazeno)-imidazole-4-carboxamide (DIC) and 900 mg/kg 5-[3,3-bis(2-chlorethyl)-1-triazeno]-imidazole-4-carboxamide (BIC) were given to pregnant Wistar rats at day 12 and the animals were killed 4 h after injection and at days 13-17 of gestation. Fetal tissues were used to determine total DNA, RNA, and protein and the data used to derive cell number and cell weight, RNA, and protein/cell. Both compounds reduced total fetal body weight, DNA, RNA, and protein but reduction of RNA by BIC was not statistically significant. These effects were observed 4 h after injection, increased with age (days 13-17), and were 3-4 times greater for DIC than BIC. By using the value of 6.2 mumug DNA/cell, cell number and per-cell values for weight, RNA, and protein, and weight: DNA, RNA:DNA, and protein:DNA ratios were computed. The per-cell values and ratios in the DIC-exposed animals were 8-44% greater and in BIC-treated animals 0-11% greater than control animals of the same gestational age. Percentage of body water was the same in the experimental and control animals. The differences in DNA, RNA, and protein are believed to be related to drug-induced growth retardation incident to total fetal DNA reduction resulting in diminished cell number.

The present status of prenatal detection of neural tube defects.

In experimentally induced myelocele in rats, efforts to find neural cells in amniotic fluid (AF) were unsuccessful. Creatine phosphokinase (CPK) and aldolase concentrations studied in serum of 118 and cerebrospinal fluid (CSF) in 9 patients with myelomeningocele showed serum CPK to be significantly elevated and more responsive to additional muscle injury than aldolase, but both enzymes appeared in lower concentrations in patients with myelomeningocele than those with infantile atrophy or cerebral palsy. In CSF, CPK, and aldolase concentrations averaged 4.2 I.U. and 2.7 S.L.U. per milliliter, respectively. Significant CPK elevation (p less than 0.001) was also found in AF from myeloschitic fetuses and maternal rat serum. Although these findings suggest that increased CPK concentration is an indicator of myelocele in rats, the technique is impractical for prenatal detection of human fetus occurs too late in gestation. This does not, however, preclude the value of CPK for detecting onset of paraparesis. In all myeloschitic human fetuses, the CSF communicates directly with AF for at least 3 to 4 weeks. This implies that CSF is probably the principal source of increased alpha-fetoprotein concentration encountered in AF of all pregnancies with NTD. When biological variables are recognized, it is evident that increased concentration of amniotic fluid alpha fetoprotein is a reliable indicator of fetuses with open myelocele and/or anenciphalus.